Polynucleotides encoding tethered interleukin-12 (il12) polypeptides and uses thereof

ABSTRACT

The present disclosure relates to polynucleotides encoding tethered interleukin-12 (IL-12) polypeptides comprising an IL-12 polypeptide and a membrane domain. The present disclosure also relates to vectors comprising the polynucleotides; host cells comprising the polynucleotides or vectors, polypeptides encoded by the polynucleotides; compositions comprising the polynucleotides, vectors, host cells, or polypeptides and a delivery agent; and uses thereof, including treatment of cancer.

RELATED APPLICATIONS

This application is a divisional of U.S. patent application Ser. No.16/613,732, filed Nov. 14, 2019, which is a 35 U.S.C. § 371 nationalstage filing of International Application No. PCT/US2018/033436, filedMay 18, 2018, which claims the benefit of U.S. Provisional ApplicationSer. No. 62/508,316, filed on May 18, 2017. The entire contents of theabove-referenced applications are incorporated herein by this reference.

REFERENCE TO SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted in XML format via USPTO Patent Center and is herebyincorporated by reference in its entirety. Said XML copy, created onJul. 28, 2022, is named MRNA_131_D01US_SeqList_ST26.xml and is 914.6killobytes in size.

BACKGROUND

Interleukin-12 (IL-12) is a pro-inflammatory cytokine that plays animportant role in innate and adaptive immunity. Gately, M K et al., AnnuRev Immunol. 16: 495-521 (1998). IL-12 functions primarily as a 70 kDaheterodimeric protein consisting of two disulfide-linked p35 and p40subunits. IL-12 p40 homodimers do exist, but other than functioning asan antagonist that binds the IL-12 receptor, they do not appear tomediate a biologic response. Id. The precursor form of the IL-12 p40subunit (NM 002187; P29460; also referred to as IL-12B, natural killercell stimulatory factor 2, cytotoxic lymphocyte maturation factor 2) is328 amino acids in length, while its mature form is 306 amino acidslong. The precursor form of the IL-12 p35 subunit (NM 000882; P29459;also referred to as IL-12A, natural killer cell stimulatory factor 1,cytotoxic lymphocyte maturation factor 1) is 219 amino acids in lengthand the mature form is 197 amino acids long. Id. The genes for the IL-12p35 and p40 subunits reside on different chromosomes and are regulatedindependently of each other. Gately, M K et al., Annu Rev Immunol. 16:495-521 (1998). Many different immune cells (e.g., dendritic cells,macrophages, monocytes, neutrophils, and B cells) produce IL-12 uponantigenic stimuli. The active IL-12 heterodimer is formed followingprotein synthesis. Id.

Due to its ability to activate both NK cells and cytotoxic T cells,IL-12 protein has been studied as a promising anti-cancer therapeuticsince 1994. See Nastala, C. L. et al., J Immunol 153: 1697-1706 (1994).But despite high expectations, early clinical studies did not yieldsatisfactory results. Lasek W. et al., Cancer Immunol Immunother 63:419-435, 424 (2014). Repeated administration of IL-12, in most patients,led to adaptive response and a progressive decline of IL-12-inducedinterferon gamma (IFNγ) levels in blood. Id. Moreover, while it wasrecognized that IL-12-induced anti-cancer activity is largely mediatedby the secondary secretion of IFNγ, the concomitant induction of IFNγalong with other cytokines (e.g., TNF-α) or chemokines (IP-10 or MIG) byIL-12 caused severe toxicity. Id.

In addition to the negative feedback and toxicity, the marginal efficacyof the IL-12 therapy in clinical settings may be caused by the strongimmunosuppressive environment in humans. Id. To minimize IFNγ toxicityand improve IL-12 efficacy, scientists tried different approaches, suchas different dose and time protocols for IL-12 therapy. See Sacco, S. etal., Blood 90: 4473-4479 (1997); Leonard, J. P. et al., Blood 90:2541-2548 (1997); Coughlin, C. M. et al., Cancer Res. 57: 2460-2467(1997); Asselin-Paturel, C. et al., Cancer 91: 113-122 (2001); andSaudemont, A. et al., Leukemia 16: 1637-1644 (2002). Nonetheless, theseapproaches have not significantly impacted patient survival. Kang, W.K., et al., Human Gene Therapy 12: 671-684 (2001).

Membrane-anchored versions of IL-12 have been studied as a means ofreducing toxicity associated with systemic administration, usingretroviral and adenoviral vectors for expression in tumor cells. SeePan, W-Y. et al., Mol. Ther. 20(5): 927-937 (2012). But, the use ofviral vectors presents a potential health risk, since the underlyingviruses can act as oncogenes and the viral vectors can be immunogenic.

Currently, a number of IL-12 clinical trials are on-going. Though thesemultiple clinical trials have been on-going for nearly 20 years sincethe first human clinical trial of IL-12 in 1996, an FDA-approved IL-12product is still not available. Thus, there is a need in the art for animproved therapeutic approach for using IL-12 to treat tumors.

SUMMARY OF THE DISCLOSURE

The present disclosure is directed to novel tethered interleukin-12(IL-12)-encoding polynucleotides (e.g., mRNAs) for use in treatingcancer.

Although IL-12 has been shown to have potent anti-tumor activity, itsclinical application is limited by severe systemic toxicity. Severalstrategies have been employed to address this limitation, which appearpromising. The present disclosure is based, at least in part, on astrategy of anchoring an IL-12 polypeptide to a cell membrane bydelivering an mRNA encoding an IL-12 polypeptide to the cell, therebygenerating a tethered IL-12 polypeptide with reduce systemicdistribution. Further, the present disclosure is based on the discoverythat tethered IL-12 polypeptides, encoded by mRNA, remain substantiallytethered to the cell surface (i.e., are not substantially released bycells expressing the mRNA encoded IL-12 polypeptide), thereby reducingsystemic distribution. It has also been discovered that mRNA encodedtethered IL-12 polypeptides retain IL-12 bioactivity. Specifically, mRNAencoding a tethered IL-12 polypeptide as described herein was shown toinduce an anti-tumor immune response, as indicated by an increase inCD8+ T cell proliferation and IFNγ secretion, along with a reduction intumor burden in vivo in both treated and non-treated (i.e., distal)tumors.

Accordingly, in one aspect, the disclosure provides a polynucleotidecomprising an open reading frame (ORF) comprising: (a) a first nucleicacid sequence encoding an Interleukin 12 p40 subunit (IL-12B), (b) asecond nucleic acid sequence encoding an Interleukin 12 p35 subunit(IL-12A), and (c) a nucleic acid sequence encoding a transmembranedomain, wherein the first nucleic acid sequence and the second nucleicacid sequence are linked by a nucleic acid sequence encoding a linker(“subunit linker”), and wherein the nucleic acid sequence encoding thetransmembrane domain is linked to the first or second nucleic acidsequence by a nucleic acid sequence encoding a linker (“transmembranedomain linker”).

In some embodiments, the first nucleic acid sequence is located at the5′ end of the subunit linker.

In some embodiments, the nucleic acid sequence encoding thetransmembrane domain is located at the 3′ end of the transmembranedomain linker.

In some embodiments, the polynucleotide further comprises a nucleic acidsequence encoding a signal peptide. In some embodiments, the nucleicacid sequence encoding the signal peptide is located at the 5′ end ofthe first nucleic acid sequence.

In some embodiments, the IL-12B has an amino acid sequence at leastabout 80%, at least about 90%, at least about 95%, at least 96%, atleast 97%, at least 98%, at least 99%, or 100% identical to amino acids23 to 328 of SEQ ID NO: 48, and wherein the amino acid sequence hasIL-12B activity.

In some embodiments, the IL-12A has an amino acid sequence at leastabout 80%, at least about 90%, at least about 95%, at least 96%, atleast 97%, at least 98%, at least 99%, or 100% identical to amino acids336 to 532 of SEQ ID NO: 48, and wherein the amino acid sequence hasIL-12A activity.

In some embodiments, the signal peptide comprises a sequence at leastabout 80%, at least about 90%, at least about 95%, at least 96%, atleast 97%, at least 98%, at least 99%, or 100% identical to amino acids1 to 22 of SEQ ID NO: 48.

In some embodiments, the subunit linker is a Gly/Ser linker. In someembodiments, the transmembrane domain linker is a Gly/Ser linker. Insome embodiments, the Gly/Ser linker comprises (GnS)_(m), wherein n is1, 2 3, 4, 5, 6, 7, 8, 9, 10, 15, or 20 and m is 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 15, or 20.

In some embodiments, the transmembrane domain is a type I transmembranedomain. In some embodiments, the transmembrane domain is a Cluster ofDifferentiation 8 (CD8) transmembrane domain or a Platelet-DerivedGrowth Factor Receptor (PDGF-R) transmembrane domain.

In some embodiments, the polynucleotide is DNA. In some embodiments, thepolynucleotide is RNA. In some embodiments, the polynucleotide is mRNA.

In some embodiments, the polynucleotide comprises at least onechemically modified nucleobase.

In some embodiments, the at least one chemically modified nucleobase isselected from the group consisting of pseudouracil (ψ),N1-methylpseudouracil(m1ψ), 2-thiouracil (s2U), 4′-thiouracil, 5-methylcytosine, 2-thio-1-methyl-1-deaza-pseudouracil,2-thio-1-methyl-pseudouracil, 2-thio-5-aza-uracil,2-thio-dihydropseudouracil, 2-thio-dihydrouracil, 2-thio-pseudouracil,4-methoxy-2-thio-pseudouracil, 4-methoxy-pseudouracil,4-thio-1-methyl-pseudouracil, 4-thio-pseudouracil, 5-aza-uracil,dihydropseudouracil, 5-methyluracil, 5-methoxyuracil, 2′-O-methyluracil, 1-methyl-pseudouracil (m1ψ), 5-methoxy-uracil (mo5U),5-methyl-cytosine (m5C), α-thio-guanine, α-thio-adenine, 5-cyano uracil,4′-thio uracil, 7-deaza-adenine, 1-methyl-adenine (m1A),2-methyl-adenine (m2A), N6-methyl-adenine (m6A), and 2,6-Diaminopurine,(I), 1-methyl-inosine (m1I), wyosine (imG), methylwyosine (mimG),7-deaza-guanine, 7-cyano-7-deaza-guanine (preQ0),7-aminomethyl-7-deaza-guanine (preQ1), 7-methyl-guanine (m7G),1-methyl-guanine (m1G), 8-oxo-guanine, 7-methyl-8-oxo-guanine, and twoor more combinations thereof.

In some embodiments, the nucleobases in the polynucleotide arechemically modified by at least about 10%, at least about 20%, at leastabout 30%, at least about 40%, at least about 50%, at least about 60%,at least about 70%, at least about 80%, at least about 90%, at least95%, at least 99%, or 100%. In some embodiments, the chemically modifiednucleobases are selected from the group consisting of uracil, adenine,cytosine, guanine, and any combination thereof.

In some embodiments, the uracils, adenines, cytosines or guanines arechemically modified by at least about 10%, at least about 20%, at leastabout 30%, at least about 40%, at least about 50%, at least about 60%,at least about 70%, at least about 80%, at least about 90%, at least95%, at least 99%, or 100%.

In some embodiments, the polynucleotide further comprises a nucleic acidsequence comprising a miRNA binding site. In some embodiments, the miRNAbinding site binds to miR-122. In some embodiments, the miRNA bindingsite binds to miR-122-3p or miR-122-5p.

In some embodiments, the polynucleotide further comprises a 5′ UTR. Insome embodiments, the 5′ UTR comprises a nucleic acid sequence at least90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of sequencesdisclosed herein.

In some embodiments, the polynucleotide further comprises a 3′ UTR. Insome embodiments, the 3′ UTR comprises a nucleic acid sequence at leastabout 90%, at least about 95%, at least 96%, at least 97%, at least 98%,at least 99%, or 100% identical to any one of sequences disclosedherein. In some embodiments, the miRNA binding site is located withinthe 3′ UTR.

In some embodiments, the 5′ UTR comprises a 5′ terminal cap.

In some embodiments, the 5′ terminal cap is a Cap0, Cap1, ARCA, inosine,N1-methyl-guanosine, 2′fluoro-guanosine, 7-deaza-guanosine,8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine, 2-azidoguanosine,Cap2, Cap4, 5′ methylG cap, or an analog thereof.

In some embodiments, the polynucleotide further comprises a poly-Aregion. In some embodiments, the poly-A region is at least about 10, atleast about 20, at least about 30, at least about 40, at least about 50,at least about 60, at least about 70, at least about 80, or at leastabout 90 nucleotides in length. In some embodiments, the poly-A regionhas about 10 to about 200 nucleotides in length, about 20 to about 180nucleotides in length, about 30 to about 160 nucleotides in length,about 40 to about 140 nucleotides in length, about 50 to about 120nucleotides in length, about 60 to about 100 nucleotides in length, orabout 80 to about 90 nucleotides in length.

In some embodiments, the polynucleotide has been transcribed in vitro(IVT). In some embodiments, the polynucleotide is chimeric. In someembodiments, the polynucleotide is circular.

In some embodiments, the ORF further comprises one or more nucleic acidsequences encoding one or more heterologous polypeptides fused to thenucleic acid sequence encoding the IL-12B, the IL-12A, or both. In someembodiments, the one or more heterologous polypeptides increase apharmacokinetic property of the IL-12A, the IL-12B, or both.

In some embodiments, the polynucleotide is single stranded. In someembodiments, the polynucleotide is double stranded.

In some embodiments, the IL-12B is a variant, derivative or mutanthaving an IL-12B activity. In some embodiments, the IL-12A is a variant,derivative, or mutant having an IL-12A activity. In another aspect, thedisclosure provides a vector comprising any of the abovepolynucleotides.

In another aspect, the disclosure provides a composition comprising (i)any of the above polynucleotides or vector, and (ii) a delivery agent.In some embodiments, the delivery agent comprises a lipid nanoparticle.In some embodiments, the lipid nanoparticle comprises the compound offormula (I). In some embodiments, the delivery agent further comprises aphospholipid. In some embodiments, the delivery agent further comprisesa structural lipid. In some embodiments, the structural lipid ischolesterol. In some embodiments, the delivery agent further comprises aPEG lipid.

In some embodiments, the delivery agent further comprises a quaternaryamine compound. In another aspect, the disclosure provides a method ofreducing the size of a tumor or inhibiting growth of a tumor in asubject in need thereof comprising administering any of the abovepolynucleotides, vector, or compositions in the subject. In someembodiments, the polynucleotide, vector, or composition is administeredsubcutaneously, intravenously, intraperitoneally, or intratumorally.

In some embodiments, the administration treats a cancer. In someembodiments, the polynucleotide is administered intratumorally to thesubject. In some embodiments, the polynucleotide is administered at anamount between about 0.10 μg per tumor and about 1000 mg per tumor.

In some embodiments, the method further comprises administering ananti-cancer agent. In some embodiments, the anti-cancer agent comprises(i) an antibody or antigen-binding fragment thereof that specificallybinds to PD-1 or PD-L1 (anti-PD-1 antibody or anti-PD-L1 antibody,respectively) or a polynucleotide encoding the anti-PD-1 or anti-PD-L1antibody or antigen-binding fragment thereof, (ii) an antibody orantigen-binding fragment thereof that specifically binds to CTLA-4(anti-CTLA-4 antibody) or a polynucleotide encoding the anti-CTLA-4antibody or antigen-binding fragment thereof, or (iii) an anti-PD-1 oranti-PD-L1 antibody or antigen-binding fragment thereof or apolynucleotide encoding the anti-PD-1 or anti-PD-LI antibody orantigen-binding fragment thereof, and an anti-CTLA-4 antibody orantigen-binding fragment thereof or a polynucleotide encoding theanti-CTLA-4 antibody or antigen-binding fragment thereof.

In some embodiments, the administration reduces the size of a tumor orinhibits growth of a tumor at least 1.5 fold, at least 2 fold, at least2.5 fold, at least 3 fold, at least 3.5 fold, at least 4 fold, at least4.5 fold, or at least 5 fold better than (i) an administration of apolynucleotide encoding IL-12 alone, (ii) an administration of theanti-PD-1 or anti-PD-L1 antibody alone, or (iii) an administration ofthe anti-CTLA-4 antibody alone.

In some embodiments, the polynucleotide encoding the anti-PD-1,anti-PD-L1, or anti-CTLA-4 antibody or antigen-binding fragment thereofcomprises an mRNA.

In some embodiments, the polynucleotide encoding the anti-PD-1,anti-PD-L1, or anti-CTLA-4 antibody or antigen-binding fragment thereofcomprises at least one chemically modified nucleoside.

In some embodiments, the at least one chemically modified nucleoside isselected from any chemically modified nucleoside disclosed herein and acombination thereof.

In some embodiments, the at least one chemically modified nucleoside isselected from the group consisting of pseudouridine,N1-methylpseudouridine, 5-methylcytosine, 5-methoxyuridine, and acombination thereof.

In some embodiments, the mRNA encoding the anti-PD-1, anti-PD-L1, oranti-CTLA-4 antibody or antigen-binding fragment thereof comprises anopen reading frame. In some embodiments, the anti-PD-L1 antibody isatezolizumab, avelumab, or durvalumab. In some embodiments, theanti-CTLA-4 antibody is tremelimumab or ipilimumab.

In some aspects, the disclosure provides a polynucleotide comprising anopen reading frame (ORF) encoding a human interleukin-12 (IL-12)polypeptide operably linked to a membrane domain, optionally via alinker, wherein the human IL-12 polypeptide comprises an IL-12 p40subunit (IL-12B) polypeptide operably linked to an IL-12 p35 subunit(IL-12A) polypeptide, wherein the membrane domain comprises atransmembrane domain, and wherein the polynucleotide is an mRNAcomprising a 5′ untranslated region (UTR), the ORF, and a 3′ UTR.

In some aspects, the disclosure provides a polynucleotide comprising anopen reading frame (ORF) encoding a human interleukin-12 (IL-12)polypeptide operably linked to a membrane domain, optionally via alinker, wherein the human IL-12 polypeptide comprises an IL-12 p40subunit (IL-12B) polypeptide operably linked to an IL-12 p35 subunit(IL-12A) polypeptide, wherein the membrane domain comprises atransmembrane domain derived from a Type I integral membrane protein,and wherein the polynucleotide is an mRNA comprising a 5′ untranslatedregion (UTR), the ORF, and a 3′ UTR.

In some aspects, the transmembrane domain is selected from the groupconsisting of: a Cluster of Differentiation 8 (CD8) transmembranedomain, a Platelet-Derived Growth Factor Receptor (PDGFR) transmembranedomain, and a Cluster of Differentiation 80 (CD80) transmembrane domain.

In some aspects, the disclosure provides a polynucleotide comprising anopen reading frame (ORF) encoding a human interleukin-12 (IL-12)polypeptide operably linked to a membrane domain, optionally via alinker, wherein the human IL-12 polypeptide comprises an IL-12 p40subunit (IL-12B) polypeptide operably linked to an IL-12 p35 subunit(IL-12A) polypeptide, wherein the membrane domain comprises a Cluster ofDifferentiation 8 (CD8) transmembrane domain, and wherein thepolynucleotide is an mRNA comprising a 5′ untranslated region (UTR), theORF, and a 3′ UTR. In some aspects, the transmembrane domain is a CD8transmembrane domain comprising the amino acid sequence set forth in SEQID NO: 101.

In some aspects, the disclosure provides a polynucleotide comprising anopen reading frame (ORF) encoding a human interleukin-12 (IL-12)polypeptide operably linked to a membrane domain, optionally via alinker, wherein the human IL-12 polypeptide comprises an IL-12 p40subunit (IL-12B) polypeptide operably linked to an IL-12 p35 subunit(IL-12A) polypeptide, wherein the membrane domain comprises aPlatelet-Derived Growth Factor Receptor (PDGFR) transmembrane domain,and wherein the polynucleotide is an mRNA comprising a 5′ untranslatedregion (UTR), the ORF, and a 3′ UTR. In some aspects, the transmembranedomain is a PDGFR transmembrane domain comprising a PDGFR-betatransmembrane domain. In some embodiments, the PDGFR-beta transmembranedomain comprises the amino acid sequence set forth in SEQ ID NO: 102.

In some aspects, the disclosure provides a polynucleotide comprising anopen reading frame (ORF) encoding a human interleukin-12 (IL-12)polypeptide operably linked to a membrane domain, optionally via alinker, wherein the human IL-12 polypeptide comprises an IL-12 p40subunit (IL-12B) polypeptide operably linked to an IL-12 p35 subunit(IL-12A) polypeptide, wherein the membrane domain comprises a Cluster ofDifferentiation 80 (CD80) transmembrane domain, and wherein thepolynucleotide is an mRNA comprising a 5′ untranslated region (UTR), theORF, and a 3′ UTR. In some aspects, the transmembrane domain is a CD80transmembrane domain comprising the amino acid sequence set forth in SEQID NO: 103.

In some aspects, the disclosure provides a polynucleotide comprising anopen reading frame (ORF) encoding a human interleukin-12 (IL-12)polypeptide operably linked to a membrane domain, optionally via alinker, wherein the human IL-12 polypeptide comprises an IL-12 p40subunit (IL-12B) polypeptide operably linked to an IL-12 p35 subunit(IL-12A) polypeptide, wherein the membrane domain comprises atransmembrane domain and an intracellular domain, and wherein thepolynucleotide is an mRNA comprising a 5′ untranslated region (UTR), theORF, and a 3′ UTR.

In some embodiments, the intracellular domain is derived from the samepolypeptide as the transmembrane domain. In some embodiments, theintracellular domain is derived from a different polypeptide than thetransmembrane domain is derived from. In some embodiments, theintracellular domain is selected from the group consisting of: a PDGFRintracellular domain, a truncated PDGFR intracellular domain, and a CD80intracellular domain.

In some aspects, the intracellular domain is a PDGFR intracellulardomain comprising a PDGFR-beta intracellular domain. In someembodiments, the PDGFR-beta intracellular domain comprises the aminoacid sequence set forth in SEQ ID NO: 226. In some aspects, thetruncated PDGFR intracellular domain comprises a PDGFR-betaintracellular domain truncated at E570 or G739. In some aspects,truncated PDGFR-beta intracellular domain truncated at E570 comprisesthe amino acid sequence set forth in SEQ ID NO: 227. In some aspects,the truncated PDGFR-beta transmembrane truncated at G739 comprises theamino acid sequence set forth in SEQ ID NO: 228.

In some aspects, the intracellular domain is a CD80 intracellulardomain. In some aspects, the CD80 intracellular domain comprises theamino acid sequence set forth in SEQ ID NO: 225.

In any of the foregoing aspects, the membrane domain comprises aPDGFR-beta transmembrane domain and a PDGFR-beta intracellular domain.In any of the foregoing aspects, the membrane domain comprises aPDGFR-beta transmembrane domain and a truncated PDGFR-beta intracellulardomain truncated at E570. In any of the foregoing aspects, the membranedomain comprises a PDGFR-beta transmembrane domain and a truncatedPDGFR-beta intracellular domain truncated at G739. In any of theforegoing aspects, the membrane domain comprises a CD80 transmembranedomain and a CD80 intracellular domain.

In some aspects, the disclosure provides a polynucleotide comprising anopen reading frame (ORF) encoding a human interleukin-12 (IL-12)polypeptide operably linked to a membrane domain, optionally via alinker, wherein the human IL-12 polypeptide comprises an IL-12 p40subunit (IL-12B) polypeptide operably linked to an IL-12 p35 subunit(IL-12A) polypeptide, wherein the membrane domain comprises aPlatelet-Derived Growth Factor Receptor (PDGFR) transmembrane domain anda PDGFR intracellular domain, and wherein the polynucleotide is an mRNAcomprising a 5′ untranslated region (UTR), the ORF, and a 3′ UTR. Insome aspects, the PDGFR transmembrane domain comprises a PDGFR-betatransmembrane domain and the PDGFR intracellular domain comprises aPDGFR-beta intracellular domain. In some aspects, the PDGFRtransmembrane domain comprises a PDGFR-beta transmembrane domain and thePDGFR intracellular domain comprises a truncated PDGFR-betaintracellular domain. In some embodiments, the PDGFR-beta transmembranedomain comprises the amino acid sequence set forth in SEQ ID NO: 102 andthe PDGFR-beta intracellular domain comprises the amino acid sequenceset forth in SEQ ID NO: 226. In some embodiments, the PDGFR-betatransmembrane domain comprises the amino acid sequence set forth in SEQID NO: 102 and the truncated PDGFR-beta intracellular domain comprisesthe amino acid sequence set forth in SEQ ID NO: 227. In someembodiments, the PDGFR-beta transmembrane domain comprises the aminoacid sequence set forth in SEQ ID NO: 102 and the truncated PDGFR-betaintracellular domain comprises the amino acid sequence set forth in SEQID NO: 228.

In some aspects, the disclosure provides a polynucleotide comprising anopen reading frame (ORF) encoding a human interleukin-12 (IL-12)polypeptide operably linked to a membrane domain, optionally via alinker, wherein the human IL-12 polypeptide comprises an IL-12 p40subunit (IL-12B) polypeptide operably linked to an IL-12 p35 subunit(IL-12A) polypeptide, wherein the membrane domain comprises a Cluster ofDifferentiation 80 (CD80) transmembrane domain and a CD80 intracellulardomain, and wherein the polynucleotide is an mRNA comprising a 5′untranslated region (UTR), the ORF, and a 3′ UTR. In some aspects, theCD80 transmembrane domain comprises the amino acid sequence set forth inSEQ ID NO: 103 and the CD80 intracellular domain comprises the aminoacid sequence set forth in SEQ ID NO: 225.

In any of the foregoing aspects, the membrane domain is operably linkedto the IL-12A polypeptide by a peptide linker. In any of the foregoingaspects, the membrane domain is operably linked to the IL-12Bpolypeptide by a peptide linker.

In some aspects, the membrane domain is operably linked to the IL-12Apolypeptide by a Gly/Ser linker. In some aspects, the membrane domain isoperably linked to the IL-12B polypeptide by a Gly/Ser linker.

In some aspects, the Gly/Ser linker comprises (G_(n)S)_(m), wherein n is1, 2 3, 4, 5, 6, 7, 8, 9, 10, 15, or 20 and m is 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 15, or 20. In some aspects, the Gly/Ser linker further comprisesa leucine and a glutamine at the 3′end of the Gly/Ser linker. In someaspects, the linker comprises the amino acid sequence set forth in SEQID NO: 229.

In any of the foregoing aspects, the IL-12B polypeptide is operablylinked to the IL-12A polypeptide by a peptide linker. In some aspects,the IL-12B polypeptide is located at the 5′ terminus of the IL-12Apolypeptide, or the 5′ terminus of the peptide linker. In some aspects,the IL-12A polypeptide is located at the 5′ terminus of the IL-12Bpolypeptide, or the 5′ terminus of the peptide linker. In some aspects,the peptide linker comprises a Gly/Ser linker. In some aspects, theGly/Ser linker comprises (G_(n)S)_(m), wherein n is 1, 2 3, 4, 5, 6, 7,8, 9, 10, 15, or 20 and m is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, or 20.In some aspects, the Gly/Ser linker comprises (G_(n)S)_(m), and whereinn is 6 and m is 1.

In any of the foregoing aspects, the ORF encodes a signal peptide. Insome aspects, the signal peptide is an IL-12B signal peptide. In someaspects, the IL-12B signal peptide comprises the amino acid sequence setforth in amino acids 1 to 22 of SEQ ID NO: 48.

In any of the foregoing aspects, the IL-12B polypeptide comprises theamino acid sequence set forth in amino acids 23 to 328 of SEQ ID NO: 48.In any of the foregoing aspects, the IL-12A polypeptide comprises theamino acid sequence set forth in amino acids 336 to 532 of SEQ ID NO:48.

In some aspects, the disclosure provides a polynucleotide comprising anopen reading frame (ORF) encoding a human interleukin-12 (IL-12)polypeptide operably linked to a membrane domain via a linker, whereinthe human IL-12 polypeptide comprises an IL-12 p40 subunit (IL-12B)polypeptide operably linked to an IL-12 p35 subunit (IL-12A) polypeptidevia a linker, wherein the membrane domain comprises a transmembranedomain, and wherein the polynucleotide is an mRNA comprising a 5′untranslated region (UTR), the ORF, and a 3′ UTR.

In some aspects, the disclosure provides a polynucleotide comprising anopen reading frame (ORF) encoding a human interleukin-12 (IL-12)polypeptide operably linked to a membrane domain via a linker, whereinthe human IL-12 polypeptide comprises an IL-12 p40 subunit (IL-12B)polypeptide operably linked to an IL-12 p35 subunit (IL-12A) polypeptidevia a linker, wherein the membrane domain comprises a transmembranedomain derived from a Type I integral membrane protein, and wherein thepolynucleotide is an mRNA comprising a 5′ untranslated region (UTR), theORF, and a 3′ UTR.

In some aspects, the transmembrane domain is selected from the groupconsisting of: a Cluster of Differentiation 8 (CD8) transmembranedomain, a Platelet-Derived Growth Factor Receptor (PDGFR) transmembranedomain, and a Cluster of Differentiation 80 (CD80) transmembrane domain.

In some aspects, the disclosure provides a polynucleotide comprising anopen reading frame (ORF) encoding a human interleukin-12 (IL-12)polypeptide operably linked to a membrane domain via a linker, whereinthe human IL-12 polypeptide comprises an IL-12 p40 subunit (IL-12B)polypeptide operably linked to an IL-12 p35 subunit (IL-12A) polypeptidevia a linker, wherein the membrane domain comprises a Cluster ofDifferentiation 8 (CD8) transmembrane domain, and wherein thepolynucleotide is an mRNA comprising a 5′ untranslated region (UTR), theORF, and a 3′ UTR. In some aspects, the transmembrane domain is a CD8transmembrane domain comprising the amino acid sequence set forth in SEQID NO: 101.

In some aspects, the disclosure provides a polynucleotide comprising anopen reading frame (ORF) encoding a human interleukin-12 (IL-12)polypeptide operably linked to a membrane domain via a linker, whereinthe human IL-12 polypeptide comprises an IL-12 p40 subunit (IL-12B)polypeptide operably linked to an IL-12 p35 subunit (IL-12A) polypeptidevia a linker, wherein the membrane domain comprises a Platelet-DerivedGrowth Factor Receptor (PDGFR) transmembrane domain, and wherein thepolynucleotide is an mRNA comprising a 5′ untranslated region (UTR), theORF, and a 3′ UTR. In some aspects, the transmembrane domain is a PDGFRtransmembrane domain comprising a PDGFR-beta transmembrane domain. Insome embodiments, the PDGFR-beta transmembrane domain comprises theamino acid sequence set forth in SEQ ID NO: 102.

In some aspects, the disclosure provides a polynucleotide comprising anopen reading frame (ORF) encoding a human interleukin-12 (IL-12)polypeptide operably linked to a membrane domain via a linker, whereinthe human IL-12 polypeptide comprises an IL-12 p40 subunit (IL-12B)polypeptide operably linked to an IL-12 p35 subunit (IL-12A) polypeptidevia a linker, wherein the membrane domain comprises a Cluster ofDifferentiation 80 (CD80) transmembrane domain, and wherein thepolynucleotide is an mRNA comprising a 5′ untranslated region (UTR), theORF, and a 3′ UTR. In some aspects, the transmembrane domain is a CD8transmembrane domain comprising the amino acid sequence set forth in SEQID NO: 103.

In some aspects, the disclosure provides a polynucleotide comprising anopen reading frame (ORF) encoding a human interleukin-12 (IL-12)polypeptide operably linked to a membrane domain via a linker, whereinthe human IL-12 polypeptide comprises an IL-12 p40 subunit (IL-12B)polypeptide operably linked to an IL-12 p35 subunit (IL-12A) polypeptidevia a linker, wherein the membrane domain comprises a Platelet-DerivedGrowth Factor Receptor (PDGFR) transmembrane domain and a PDGFRintracellular domain, and wherein the polynucleotide is an mRNAcomprising a 5′ untranslated region (UTR), the ORF, and a 3′ UTR. Insome aspects, the PDGFR transmembrane domain comprises a PDGFR-betatransmembrane domain and the PDGFR intracellular domain comprises aPDGFR-beta intracellular domain. In some aspects, the PDGFRtransmembrane domain comprises a PDGFR-beta transmembrane domain and thePDGFR intracellular domain comprises a truncated PDGFR-betaintracellular domain. In some embodiments, the PDGFR-beta transmembranedomain comprises the amino acid sequence set forth in SEQ ID NO: 102 andthe PDGFR-beta intracellular domain comprises the amino acid sequenceset forth in SEQ ID NO: 226. In some embodiments, the PDGFR-betatransmembrane domain comprises the amino acid sequence set forth in SEQID NO: 102 and the truncated PDGFR-beta intracellular domain comprisesthe amino acid sequence set forth in SEQ ID NO: 227. In someembodiments, the PDGFR-beta transmembrane domain comprises the aminoacid sequence set forth in SEQ ID NO: 102 and the truncated PDGFR-betaintracellular domain comprises the amino acid sequence set forth in SEQID NO: 228.

In some aspects, the disclosure provides a polynucleotide comprising anopen reading frame (ORF) encoding a human interleukin-12 (IL-12)polypeptide operably linked to a membrane domain via a linker, whereinthe human IL-12 polypeptide comprises an IL-12 p40 subunit (IL-12B)polypeptide operably linked to an IL-12 p35 subunit (IL-12A) polypeptidevia a linker, wherein the membrane domain comprises a Cluster ofDifferentiation 80 (CD80) transmembrane domain and a CD80 intracellulardomain, and wherein the polynucleotide is an mRNA comprising a 5′untranslated region (UTR), the ORF, and a 3′ UTR. In some aspects, theCD80 transmembrane domain comprises the amino acid sequence set forth inSEQ ID NO: 103 and the CD80 intracellular domain comprises the aminoacid sequence set forth in SEQ ID NO: 225.

In some aspects, the disclosure provides an mRNA comprising a 5′ UTR, anopen reading frame (ORF), and a 3′ UTR, wherein the ORF comprises anucleotide sequence encoding from 5′ to 3′:

-   -   5′-[IL-12B]-[L1]-[IL-12A]-[L2]-[MD]-3′, wherein

IL-12B is a human IL-12 p40 subunit polypeptide,

L1 is a first peptide linker,

IL-12A is a human IL-12 p35 subunit polypeptide,

L2 is a second peptide linker,

MD is a membrane domain comprising a transmembrane domain from a Type Iintegral membrane protein. In some aspects, the Type I integral membraneprotein is selected from the group consisting of: a Cluster ofDifferentiation 8 (CD8) transmembrane domain, a Platelet-Derived GrowthFactor Receptor (PDGFR) transmembrane domain, and a Cluster ofDifferentiation 80 (CD80) transmembrane domain.

In some aspects, the disclosure provides an mRNA comprising a 5′ UTR, anopen reading frame (ORF), and a 3′ UTR, wherein the ORF comprises anucleotide sequence encoding from 5′ to 3′:

5′-[IL-12B]-[L1]-[IL-12A]-[L2]-[MD]-3′, wherein

IL-12B is a human IL-12 p40 subunit polypeptide,

L1 is a first peptide linker,

IL-12A is a human IL-12 p35 subunit polypeptide,

L2 is a second peptide linker,

MD is a membrane domain comprising a transmembrane domain from a Type Iintegral membrane protein,

wherein the Type I integral membrane protein is selected from the groupconsisting of: a Cluster of Differentiation 8 (CD8) transmembranedomain, a Platelet-Derived Growth Factor Receptor (PDGFR) transmembranedomain, and a Cluster of Differentiation 80 (CD80) transmembrane domain.

In some aspects, the disclosure provides an mRNA comprising a 5′ UTR, anopen reading frame (ORF), and a 3′ UTR, wherein the ORF comprises anucleotide sequence encoding from 5′ to 3′:

-   -   5′-[IL-12B]-[L1]-[IL-12A]-[L2]-[MD]-3′, wherein

IL-12B is a human IL-12 p40 subunit polypeptide,

L1 is a first peptide linker,

IL-12A is a human IL-12 p35 subunit polypeptide,

L2 is a second peptide linker,

MD is a membrane domain comprising a transmembrane domain from a Type Iintegral membrane protein and an intracellular domain. In some aspects,the Type I integral membrane protein is selected from the groupconsisting of: a Cluster of Differentiation 8 (CD8) transmembranedomain, a Platelet-Derived Growth Factor Receptor (PDGFR) transmembranedomain, and a Cluster of Differentiation 80 (CD80) transmembrane domain.

In some aspects, the disclosure provides an mRNA comprising a 5′ UTR, anopen reading frame (ORF), and a 3′ UTR, wherein the ORF comprises anucleotide sequence encoding from 5′ to 3′:

-   -   5′-[IL-12B]-[L1]-[IL-12A]-[L2]-[MD]-3′, wherein

IL-12B is a human IL-12 p40 subunit polypeptide,

L1 is a first peptide linker,

IL-12A is a human IL-12 p35 subunit polypeptide,

L2 is a second peptide linker,

MD is a membrane domain comprising a transmembrane domain from a Type Iintegral membrane protein and an intracellular domain,

wherein the Type I integral membrane protein is selected from the groupconsisting of: a Cluster of Differentiation 8 (CD8) transmembranedomain, a Platelet-Derived Growth Factor Receptor (PDGFR) transmembranedomain, and a Cluster of Differentiation 80 (CD80) transmembrane domain.

In some aspects, the disclosure provides an mRNA comprising a 5′ UTR, anopen reading frame (ORF), and a 3′ UTR, wherein the ORF comprises anucleotide sequence encoding from 5′ to 3′:

-   -   5′-[IL-12B]-[L1]-[IL-12A]-[L2]-[MD]-3′, wherein

IL-12B is a human IL-12 p40 subunit polypeptide,

L1 is a first peptide linker,

IL-12A is a human IL-12 p35 subunit polypeptide,

L2 is a second peptide linker,

MD is a membrane domain comprising a Cluster of Differentiation Factor80 (CD80) transmembrane domain. In some aspects, the CD80 transmembranedomain comprises the amino acid sequence set forth in SEQ ID NO: 103.

In some aspects, the disclosure provides an mRNA comprising a 5′ UTR, anopen reading frame (ORF), and a 3′ UTR, wherein the ORF comprises anucleotide sequence encoding from 5′ to 3′:

-   -   5′-[IL-12B]-[L1]-[IL-12A]-[L2]-[MD]-3′, wherein

IL-12B is a human IL-12 p40 subunit polypeptide,

L1 is a first peptide linker,

IL-12A is a human IL-12 p35 subunit polypeptide,

L2 is a second peptide linker,

MD is a membrane domain comprising a Cluster of Differentiation Factor80 (CD80) transmembrane domain and a CD80 intracellular domain. In someaspects, the CD80 transmembrane domain comprises the amino acid sequenceset forth in SEQ ID NO: 103. In some aspects, the CD80 intracellulardomain comprises the amino acid sequence set forth in SEQ ID NO: 225. Insome aspects, the MD comprises a CD80 transmembrane domain comprisingthe amino acid sequence set forth in SEQ ID NO: 103, and a CD80intracellular domain comprising the amino acid sequence set forth in SEQID NO: 225.

In some aspects, the disclosure provides an mRNA comprising a 5′ UTR, anopen reading frame (ORF), and a 3′ UTR, wherein the ORF comprises anucleotide sequence encoding from 5′ to 3′:

-   -   5′-[IL-12B]-[L1]-[IL-12A]-[L2]-[MD]-3′, wherein

IL-12B is a human IL-12 p40 subunit polypeptide,

L1 is a first peptide linker,

IL-12A is a human IL-12 p35 subunit polypeptide,

L2 is a second peptide linker,

MD is a membrane domain comprising a PDGFR transmembrane domain. In someaspects, the PDGFR transmembrane domain comprises a PDGFR-betatransmembrane domain. In some aspects, the PDGFR-beta transmembranedomain comprises the amino acid sequence set forth in SEQ ID NO: 102.

In some aspects, the disclosure provides an mRNA comprising a 5′ UTR, anopen reading frame (ORF), and a 3′ UTR, wherein the ORF comprises anucleotide sequence encoding from 5′ to 3′:

-   -   5′-[IL-12B]-[L1]-[IL-12A]-[L2]-[MD]-3′, wherein

IL-12B is a human IL-12 p40 subunit polypeptide,

L1 is a first peptide linker,

IL-12A is a human IL-12 p35 subunit polypeptide,

L2 is a second peptide linker,

MD is a membrane domain comprising a PDGFR transmembrane domain and aPDGFR intracellular domain. In some aspects, the PDGFR transmembranedomain comprises a PDGFR-beta transmembrane domain. In some aspects, thePDGFR-beta transmembrane domain comprises the amino acid sequence setforth in SEQ ID NO: 102. In some aspects, the PDGFR intracellular domaincomprises a PDGFR-beta intracellular domain. In some aspects, thePDGFR-beta intracellular domain comprises the amino acid sequence setforth in SEQ ID NO: 226. In some aspects, the PDGFR intracellular domaincomprises a truncated PDGFR-beta intracellular domain. In some aspects,the truncated PDGFR-beta intracellular domain is truncated at E570 andcomprises the amino acid sequence set forth in SEQ ID NO: 227. In someaspects, the truncated PDGFR-beta intracellular domain is truncated atG739 and comprises the amino acid sequence set forth in SEQ ID NO: 228.In some aspects, the MD comprises a PDGFR-beta transmembrane domaincomprising the amino acid sequence set forth in SEQ ID NO: 102, and aPDGFR-beta intracellular domain comprising the amino acid sequence setforth in SEQ ID NO: 226. In some aspects, the MD comprises a PDGFR-betatransmembrane domain comprising the amino acid sequence set forth in SEQID NO: 102, and a truncated PDGFR-beta intracellular domain comprisingthe amino acid sequence set forth in SEQ ID NO: 227. In some aspects,the MD comprises a PDGFR-beta transmembrane domain comprising the aminoacid sequence set forth in SEQ ID NO: 102, and a truncated PDGFR-betaintracellular domain comprising the amino acid sequence set forth in SEQID NO: 228.

In any of the foregoing aspects, the ORF of the mRNA encodes a signalpeptide. In some aspects, the signal peptide is an IL-12B signalpeptide. In some aspects, the IL-12B signal peptide comprises the aminoacid sequence set forth in amino acids 1 to 22 of SEQ ID NO: 48.

In any of the foregoing aspects, the ORF of the mRNA encodes an IL-12Bpolypeptide comprising the amino acid sequence set forth in amino acids23 to 328 of SEQ ID NO: 48.

In any of the foregoing aspects, the ORF of the mRNA encodes an IL-12Bpolypeptide comprising the amino acid sequence set forth in amino acids336 to 532 of SEQ ID NO: 48.

In any of the foregoing aspects, first peptide linker [L1] and secondpeptide linker [L2] are each a Gly/Ser linker. In some aspects, [L1]comprises SEQ ID NO: 214. In some aspects, [L2] comprises (G_(n)S)_(m),wherein n is 1-4, 1, 2, 3 or 4 and m is 1-4, 1, 2, 3, or 4. In someaspects, [L2] comprises (G₄S)_(m), wherein m is 1-4, 1, 2, 3, or 4. Insome aspects, [L2] comprises the amino acid sequence set forth in SEQ IDNO: 229.

In any of the foregoing aspects, the ORF of the mRNA comprises thesequence set forth in SEQ ID NO: 273 or SEQ ID NO: 274, or a nucleotidesequence at least 90%, at least 95%, at least 97%, at least 98% or atleast 99% identical to a sequence set forth in SEQ ID NO: 273 or SEQ IDNO: 274.

In any of the foregoing aspects, the ORF of the mRNA comprises thesequence set forth in any one of SEQ ID NOs: 275-279, or a nucleotidesequence at least 90%, at least 95%, at least 97%, at least 98% or atleast 99% identical to a sequence set forth in any one of SEQ ID NOs:275-279.

In any of the foregoing aspects, the ORF of the mRNA comprises thesequence set forth in SEQ ID NO: 281, or a nucleotide sequence at least90%, at least 95%, at least 97%, at least 98% or at least 99% identicalto a sequence set forth in SEQ ID NO: 281.

In any of the foregoing aspects, the ORF of the mRNA comprises thesequence set forth in SEQ ID NO: 282, or a nucleotide sequence at least90%, at least 95%, at least 97%, at least 98% or at least 99% identicalto a sequence set forth in SEQ ID NO: 282.

In any of the foregoing aspects, the 3′UTR of the polynucleotide or mRNAcomprises a microRNA binding site. In some aspects, the microRNA bindingsite is a miR-122 binding site. In some aspects, the miR-122 bindingsite is a miR-122-3p or miR-122-5p binding site. In some aspects, themiR-122-5p binding site comprises the sequence set forth in SEQ ID NO:54. In some aspects, the 3′UTR comprises a sequence set forth in SEQ IDNO: 283.

In any of the foregoing aspects, the 5′UTR of the polynucleotide or mRNAcomprises a sequence set forth in SEQ ID NO: 287.

In any of the foregoing aspects, the polynucleotide or mRNA comprises a5′terminal cap structure. In some aspects, the 5′ terminal cap structureis a Cap0, Cap1, ARCA, inosine, N1-methyl-guanosine, 2′fluoro-guanosine,7-deaza-guanosine, 8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine,2-azidoguanosine, Cap2, Cap4, 5′methylGcap, or an analog thereof.

In any of the foregoing aspects, the polynucleotide or mRNA comprises a3′ polyA tail.

In any of the foregoing aspects, the polynucleotide or mRNA comprises atleast one chemical modification. In some aspects, the chemicalmodification is selected from the group consisting of pseudouridine,N1-methylpseudouridine, 2-thiouridine, 4′-thiouridine, 5-methylcytosine,2-thio-1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-pseudouridine,2-thio-5-aza-uridine, 2-thio-dihydropseudouridine,2-thio-dihydrouridine, 2-thio-pseudouridine,4-methoxy-2-thio-pseudouridine, 4-methoxy-pseudouridine,4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine,dihydropseudouridine, 5-methyluridine, 5-methyluridine,5-methoxyuridine, and 2′-O-methyl uridine. In some aspects, the chemicalmodification is selected from the group consisting of pseudouridine or apseudouridine analog. In some aspects, the chemical modification isN1-methylpseudouridine. In some aspects, the mRNA is fully modified withN1-methylpseudouridine.

In some aspects, the disclosure provides a composition comprising apolynucleotide or mRNA as described herein, and a pharmaceuticallyacceptable carrier.

In some aspects, the disclosure provides a lipid nanoparticle comprisinga polynucleotide or mRNA as described herein.

In some aspects, the disclosure provides a lipid nanoparticle comprisinga polynucleotide comprising an open reading frame (ORF) encoding a humaninterleukin-12 (IL-12) polypeptide operably linked to a membrane domain,optionally via a linker, wherein the human IL-12 polypeptide comprisesan IL-12 p40 subunit (IL-12B) polypeptide operably linked to an IL-12p35 subunit (IL-12A) polypeptide, wherein the membrane domain comprisesa transmembrane domain, and wherein the polynucleotide is an mRNAcomprising a 5′ untranslated region (UTR), the ORF, and a 3′ UTR.

In some aspects, the disclosure provides a lipid nanoparticle comprisinga polynucleotide comprising an open reading frame (ORF) encoding a humaninterleukin-12 (IL-12) polypeptide operably linked to a membrane domain,via a linker, wherein the human IL-12 polypeptide comprises an IL-12 p40subunit (IL-12B) polypeptide operably linked to an IL-12 p35 subunit(IL-12A) polypeptide, wherein the membrane domain comprises atransmembrane domain and an intracellular domain, and wherein thepolynucleotide is an mRNA comprising a 5′ untranslated region (UTR), theORF, and a 3′ UTR.

In some aspects, the disclosure provides a lipid nanoparticle comprisinga polynucleotide comprising an open reading frame (ORF) encoding a humaninterleukin-12 (IL-12) polypeptide operably linked to a membrane domain,via a linker, wherein the human IL-12 polypeptide comprises an IL-12 p40subunit (IL-12B) polypeptide operably linked to an IL-12 p35 subunit(IL-12A) polypeptide, wherein the membrane domain comprises a Cluster ofDifferentiation 8 (CD8) transmembrane domain, and wherein thepolynucleotide is an mRNA comprising a 5′ untranslated region (UTR), theORF, and a 3′ UTR.

In some aspects, the disclosure provides a lipid nanoparticle comprisinga polynucleotide comprising an open reading frame (ORF) encoding a humaninterleukin-12 (IL-12) polypeptide operably linked to a membrane domain,via a linker, wherein the human IL-12 polypeptide comprises an IL-12 p40subunit (IL-12B) polypeptide operably linked to an IL-12 p35 subunit(IL-12A) polypeptide, wherein the membrane domain comprises aPlatelet-Derived Growth Factor Receptor (PDGFR) transmembrane domain,and wherein the polynucleotide is an mRNA comprising a 5′ untranslatedregion (UTR), the ORF, and a 3′ UTR.

In some aspects, the disclosure provides a lipid nanoparticle comprisinga polynucleotide comprising an open reading frame (ORF) encoding a humaninterleukin-12 (IL-12) polypeptide operably linked to a membrane domain,via a linker, wherein the human IL-12 polypeptide comprises an IL-12 p40subunit (IL-12B) polypeptide operably linked to an IL-12 p35 subunit(IL-12A) polypeptide, wherein the membrane domain comprises a Cluster ofDifferentiation 80 (CD80) transmembrane domain, and wherein thepolynucleotide is an mRNA comprising a 5′ untranslated region (UTR), theORF, and a 3′ UTR.

In some aspects, the disclosure provides a lipid nanoparticle comprisinga polynucleotide comprising an open reading frame (ORF) encoding a humaninterleukin-12 (IL-12) polypeptide operably linked to a membrane domain,optionally via a linker, wherein the human IL-12 polypeptide comprisesan IL-12 p40 subunit (IL-12B) polypeptide operably linked to an IL-12p35 subunit (IL-12A) polypeptide, wherein the membrane domain comprisesa Platelet-Derived Growth Factor Receptor (PDGFR) transmembrane domainand a PDGFR intracellular domain, and wherein the polynucleotide is anmRNA comprising a 5′ untranslated region (UTR), the ORF, and a 3′ UTR.

In some aspects, the disclosure provides a lipid nanoparticle comprisinga polynucleotide comprising an open reading frame (ORF) encoding a humaninterleukin-12 (IL-12) polypeptide operably linked to a membrane domain,optionally via a linker, wherein the human IL-12 polypeptide comprisesan IL-12 p40 subunit (IL-12B) polypeptide operably linked to an IL-12p35 subunit (IL-12A) polypeptide, wherein the membrane domain comprisesa Cluster of Differentiation 80 (CD80) transmembrane domain and a CD80intracellular domain, and wherein the polynucleotide is an mRNAcomprising a 5′ untranslated region (UTR), the ORF, and a 3′ UTR.

In some aspects, the lipid nanoparticle comprises a molar ratio of about20-60% ionizable amino lipid: 5-25% phospholipid: 25-55% sterol; and0.5-15% PEG-modified lipid. In some aspects, the ionizable amino lipidis selected from the group consisting of for example,2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA),dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), anddi((Z)-non-2-en-1-yl)9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319). In someaspects, the ionizable amino lipid comprises a compound of Formula (I).In some aspects, the compound of Formula (I) is Compound 18.

In some aspects, the disclosure provides a lipid nanoparticle comprisinga polynucleotide comprising an open reading frame (ORF) encoding a humaninterleukin-12 (IL-12) polypeptide operably linked to a membrane domain,optionally via a linker, wherein the human IL-12 polypeptide comprisesan IL-12 p40 subunit (IL-12B) polypeptide operably linked to an IL-12p35 subunit (IL-12A) polypeptide, wherein the membrane domain comprisesa Cluster of Differentiation 80 (CD80) transmembrane domain and a CD80intracellular domain, wherein the polynucleotide is an mRNA comprising a5′ untranslated region (UTR), the ORF, and a 3′ UTR, and wherein thelipid nanoparticle comprising a molar raiot of about 20-60% ionizableamino lipid: 5-25% phospholipid: 25-55% sterol; and 0.5-15% PEG-modifiedlipid.

In some aspects, the disclosure provides a lipid nanoparticle comprisinga polynucleotide comprising an open reading frame (ORF) encoding a humaninterleukin-12 (IL-12) polypeptide operably linked to a membrane domain,optionally via a linker, wherein the human IL-12 polypeptide comprisesan IL-12 p40 subunit (IL-12B) polypeptide operably linked to an IL-12p35 subunit (IL-12A) polypeptide, wherein the membrane domain comprisesa Cluster of Differentiation 80 (CD80) transmembrane domain and a CD80intracellular domain, wherein the polynucleotide is an mRNA comprising a5′ untranslated region (UTR), the ORF, and a 3′ UTR, wherein the lipidnanoparticle comprising a molar raiot of about 20-60% ionizable aminolipid: 5-25% phospholipid: 25-55% sterol; and 0.5-15% PEG-modifiedlipid, and wherein the ionizable amino lipid is Compound 18.

In some aspects, the disclosure provides a pharmaceutical compositioncomprising a lipid nanoparticle as described herein, and apharmaceutically acceptable carrier. In some aspects, thepharmaceutically acceptable carrier comprises a buffer solution. In someaspects, the pharmaceutical composition is formulated for intratumoraldelivery.

In some aspects, the disclosure provides a polynucleotide, mRNA,composition, lipid nanoparticle, or pharmaceutical composition asdescribed herein, for use in treating or delaying progression of cancerin an individual, wherein the treatment comprises administration of thepolynucleotide, mRNA, composition, lipid nanoparticle or pharmaceuticalcomposition in combination with a second composition, wherein the secondcomposition comprises anti-cancer agent, or a polynucleotide comprisingan ORF encoding an anti-cancer agent, such as a checkpoint inhibitorpolypeptide, and an optional pharmaceutically acceptable carrier.

In some aspects, the disclosure provides a lipid nanoparticle comprisinga polynucleotide comprising an open reading frame (ORF) encoding a humaninterleukin-12 (IL-12) polypeptide operably linked to a membrane domain,optionally via a linker, wherein the human IL-12 polypeptide comprisesan IL-12 p40 subunit (IL-12B) polypeptide operably linked to an IL-12p35 subunit (IL-12A) polypeptide, wherein the membrane domain comprisesa transmembrane domain, and wherein the polynucleotide is an mRNAcomprising a 5′ untranslated region (UTR), the ORF, and a 3′ UTR, foruse in treating or delaying progression of cancer in an individual,wherein the treatment comprises administration of the lipid nanoparticlein combination with a second composition, wherein the second compositioncomprises anti-cancer agent, or a polynucleotide comprising an ORFencoding an anti-cancer agent, such as a checkpoint inhibitorpolypeptide, and an optional pharmaceutically acceptable carrier.

In some aspects, the disclosure provides use of a polynucleotide, mRNA,composition, lipid nanoparticle, or pharmaceutical composition describedherein in the manufacture of a medicament for treating or delayingprogression of cancer in an individual, wherein the medicament comprisesthe polynucleotide, mRNA, composition, lipid nanoparticle, orpharmaceutical composition, and an optional pharmaceutically acceptablecarrier, and wherein the treatment comprises administration of themedicament in combination with a composition comprising an anti-canceragent, or a polynucleotide comprising an ORF encoding an anti-canceragent, such as a checkpoint inhibitor polypeptide, and an optionalpharmaceutically acceptable carrier.

In some aspects, the disclosure provides use of a a lipid nanoparticlecomprising a polynucleotide comprising an open reading frame (ORF)encoding a human interleukin-12 (IL-12) polypeptide operably linked to amembrane domain, optionally via a linker, wherein the human IL-12polypeptide comprises an IL-12 p40 subunit (IL-12B) polypeptide operablylinked to an IL-12 p35 subunit (IL-12A) polypeptide, wherein themembrane domain comprises a transmembrane domain, and wherein thepolynucleotide is an mRNA comprising a 5′ untranslated region (UTR), theORF, and a 3′ UTR, in the manufacture of a medicament for treating ordelaying progression of cancer in an individual, wherein the medicamentcomprises the lipid nanoparticle and an optional pharmaceuticallyacceptable carrier, and wherein the treatment comprises administrationof the medicament in combination with a composition comprising ananti-cancer agent, or a polynucleotide comprising an ORF encoding ananti-cancer agent, such as a checkpoint inhibitor polypeptide, and anoptional pharmaceutically acceptable carrier.

In some aspects, the disclosure provides a kit comprising a containercomprising a polynucleotide, mRNA, composition, the lipid nanoparticle,or pharmaceutical composition as described herein, and an optionalpharmaceutically acceptable carrier, and a package insert comprisinginstructions for administration of the polynucleotide, mRNA,composition, lipid nanoparticle or pharmaceutical composition, fortreating or delaying progression of cancer in an individual. In someaspects, the package insert further comprises instructions foradministration of the lipid nanoparticle or pharmaceutical compositionin combination with a composition comprising an anti-cancer agent, or apolynucleotide comprising an ORF encoding an anti-cancer agent, such asa checkpoint inhibitor polypeptide, and an optional pharmaceuticallyacceptable carrier, for treating or delaying progression of cancer in anindividual.

In some aspects, the disclosure provides a kit comprising a containercomprising a lipid nanoparticle comprising a polynucleotide comprisingan open reading frame (ORF) encoding a human interleukin-12 (IL-12)polypeptide operably linked to a membrane domain, optionally via alinker, wherein the human IL-12 polypeptide comprises an IL-12 p40subunit (IL-12B) polypeptide operably linked to an IL-12 p35 subunit(IL-12A) polypeptide, wherein the membrane domain comprises atransmembrane domain, and wherein the polynucleotide is an mRNAcomprising a 5′ untranslated region (UTR), the ORF, and a 3′ UTR, and anoptional pharmaceutically acceptable carrier, and a package insertcomprising instructions for administration of the lipid nanoparticle ortreating or delaying progression of cancer in an individual.

In some aspects, the disclosure provides a kit comprising apolynucleotide, mRNA, composition, the lipid nanoparticle, orpharmaceutical composition as described herein, and a package insertcomprising instructions for administration of the medicament alone, orin combination with a composition comprising an anti-cancer agent, or apolynucleotide comprising an ORF encoding an anti-cancer agent, such asa checkpoint inhibitor polypeptide, and an optional pharmaceuticallyacceptable carrier, for treating or delaying progression of cancer in anindividual. In some aspects, the kit further comprises a package insertcomprising instructions for administration of the first medicament priorto, current with, or subsequent to administration of the secondmedicament for treating or delaying progression of cancer in anindividual.

In some aspects, the disclosure provides a kit comprising a lipidnanoparticle comprising a polynucleotide comprising an open readingframe (ORF) encoding a human interleukin-12 (IL-12) polypeptide operablylinked to a membrane domain, optionally via a linker, wherein the humanIL-12 polypeptide comprises an IL-12 p40 subunit (IL-12B) polypeptideoperably linked to an IL-12 p35 subunit (IL-12A) polypeptide, whereinthe membrane domain comprises a transmembrane domain, and wherein thepolynucleotide is an mRNA comprising a 5′ untranslated region (UTR), theORF, and a 3′ UTR, and a package insert comprising instructions foradministration of the medicament alone, or in combination with acomposition comprising an anti-cancer agent, or a polynucleotidecomprising an ORF encoding an anti-cancer agent, such as a checkpointinhibitor polypeptide, and an optional pharmaceutically acceptablecarrier, for treating or delaying progression of cancer in anindividual.

In any of the foregoing aspects, the checkpoint inhibitor polypeptideinhibits PD1, PD-L1, CTLA4, or a combination thereof. In some aspects,the checkpoint inhibitor polypeptide is an antibody. In some aspects,the checkpoint inhibitor polypeptide is an antibody selected from ananti-CTLA4 antibody or antigen-binding fragment thereof thatspecifically binds CTLA4, an anti-PD1 antibody or antigen-bindingfragment thereof that specifically binds PD1, an anti-PD-L1 antibody orantigen-binding fragment thereof that specifically binds PD-L1, and acombination thereof. In some aspects, the checkpoint inhibitorpolypeptide is an anti-PD-L1 antibody selected from atezolizumab,avelumab, or durvalumab. In some aspects, the checkpoint inhibitorpolypeptide is an anti-CTLA-4 antibody selected from tremelimumab oripilimumab. In some aspects, the checkpoint inhibitor polypeptide is ananti-PD1 antibody selected from nivolumab or pembrolizumab.

In some aspects, the disclosure provides a method of reducing the sizeof a tumor or inhibiting growth of a tumor in a subject in need thereof,comprising administering to the subject an effective amount of apolynucleotide, mRNA, composition, lipid nanoparticle or pharmaceuticalcomposition as described herein.

In some aspects, the disclosure provides a method of reducing the sizeof a tumor or inhibiting growth of a tumor in a subject in need thereof,comprising administering to the subject an effective amount of a lipidnanoparticle comprising a polynucleotide comprising an open readingframe (ORF) encoding a human interleukin-12 (IL-12) polypeptide operablylinked to a membrane domain, optionally via a linker, wherein the humanIL-12 polypeptide comprises an IL-12 p40 subunit (IL-12B) polypeptideoperably linked to an IL-12 p35 subunit (IL-12A) polypeptide, whereinthe membrane domain comprises a transmembrane domain, and wherein thepolynucleotide is an mRNA comprising a 5′ untranslated region (UTR), theORF, and a 3′ UTR.

In some aspects, the disclosure provides a method of reducing the sizeof a tumor or inhibiting growth of a tumor in a subject in need thereof,comprising administering to the subject an effective amount of a lipidnanoparticle comprising a polynucleotide comprising an open readingframe (ORF) encoding a human interleukin-12 (IL-12) polypeptide operablylinked to a membrane domain, via a linker, wherein the human IL-12polypeptide comprises an IL-12 p40 subunit (IL-12B) polypeptide operablylinked to an IL-12 p35 subunit (IL-12A) polypeptide, wherein themembrane domain comprises a transmembrane domain and an intracellulardomain, and wherein the polynucleotide is an mRNA comprising a 5′untranslated region (UTR), the ORF, and a 3′ UTR.

In some aspects, the disclosure provides a method of reducing the sizeof a tumor or inhibiting growth of a tumor in a subject in need thereof,comprising administering to the subject an effective amount of a lipidnanoparticle comprising a polynucleotide comprising an open readingframe (ORF) encoding a human interleukin-12 (IL-12) polypeptide operablylinked to a membrane domain, via a linker, wherein the human IL-12polypeptide comprises an IL-12 p40 subunit (IL-12B) polypeptide operablylinked to an IL-12 p35 subunit (IL-12A) polypeptide, wherein themembrane domain comprises a CD80 transmembrane domain, and wherein thepolynucleotide is an mRNA comprising a 5′ untranslated region (UTR), theORF, and a 3′ UTR.

In some aspects, the disclosure provides a method of reducing the sizeof a tumor or inhibiting growth of a tumor in a subject in need thereof,comprising administering to the subject an effective amount of a lipidnanoparticle comprising a polynucleotide comprising an open readingframe (ORF) encoding a human interleukin-12 (IL-12) polypeptide operablylinked to a membrane domain, via a linker, wherein the human IL-12polypeptide comprises an IL-12 p40 subunit (IL-12B) polypeptide operablylinked to an IL-12 p35 subunit (IL-12A) polypeptide, wherein themembrane domain comprises a PDGFR transmembrane domain, and wherein thepolynucleotide is an mRNA comprising a 5′ untranslated region (UTR), theORF, and a 3′ UTR.

In some aspects, the disclosure provides a method of reducing the sizeof a tumor or inhibiting growth of a tumor in a subject in need thereof,comprising administering to the subject an effective amount of a lipidnanoparticle comprising a polynucleotide comprising an open readingframe (ORF) encoding a human interleukin-12 (IL-12) polypeptide operablylinked to a membrane domain, via a linker, wherein the human IL-12polypeptide comprises an IL-12 p40 subunit (IL-12B) polypeptide operablylinked to an IL-12 p35 subunit (IL-12A) polypeptide, wherein themembrane domain comprises a CD8 transmembrane domain, and wherein thepolynucleotide is an mRNA comprising a 5′ untranslated region (UTR), theORF, and a 3′ UTR.

In some aspects, the disclosure provides a method of reducing the sizeof a tumor or inhibiting growth of a tumor in a subject in need thereof,comprising administering to the subject an effective amount of a lipidnanoparticle comprising a polynucleotide comprising an open readingframe (ORF) encoding a human interleukin-12 (IL-12) polypeptide operablylinked to a membrane domain, via a linker, wherein the human IL-12polypeptide comprises an IL-12 p40 subunit (IL-12B) polypeptide operablylinked to an IL-12 p35 subunit (IL-12A) polypeptide, wherein themembrane domain comprises a PDGFR transmembrane domain and a PDGFRintracellular domain, and wherein the polynucleotide is an mRNAcomprising a 5′ untranslated region (UTR), the ORF, and a 3′ UTR.

In some aspects, the disclosure provides a method of reducing the sizeof a tumor or inhibiting growth of a tumor in a subject in need thereof,comprising administering to the subject an effective amount of a lipidnanoparticle comprising a polynucleotide comprising an open readingframe (ORF) encoding a human interleukin-12 (IL-12) polypeptide operablylinked to a membrane domain, via a linker, wherein the human IL-12polypeptide comprises an IL-12 p40 subunit (IL-12B) polypeptide operablylinked to an IL-12 p35 subunit (IL-12A) polypeptide, wherein themembrane domain comprises a CD80 transmembrane domain and a CD80intracellular domain, and wherein the polynucleotide is an mRNAcomprising a 5′ untranslated region (UTR), the ORF, and a 3′ UTR.

In some aspects, the disclosure provides a method of inducing ananti-tumor response in a subject in need thereof, comprisingadministering to the subject an effective amount of a polynucleotide,mRNA, composition, lipid nanoparticle or pharmaceutical composition asdescribed herein.

In some aspects, the disclosure provides a method of inducing ananti-tumor response in a subject in need thereof, comprisingadministering to the subject an effective amount of a lipid nanoparticlecomprising a polynucleotide comprising an open reading frame (ORF)encoding a human interleukin-12 (IL-12) polypeptide operably linked to amembrane domain, optionally via a linker, wherein the human IL-12polypeptide comprises an IL-12 p40 subunit (IL-12B) polypeptide operablylinked to an IL-12 p35 subunit (IL-12A) polypeptide, wherein themembrane domain comprises a transmembrane domain, and wherein thepolynucleotide is an mRNA comprising a 5′ untranslated region (UTR), theORF, and a 3′ UTR.

In some aspects, the disclosure provides a method of inducing ananti-tumor response in a subject in need thereof, comprisingadministering to the subject an effective amount of a lipid nanoparticlecomprising a polynucleotide comprising an open reading frame (ORF)encoding a human interleukin-12 (IL-12) polypeptide operably linked to amembrane domain, via a linker, wherein the human IL-12 polypeptidecomprises an IL-12 p40 subunit (IL-12B) polypeptide operably linked toan IL-12 p35 subunit (IL-12A) polypeptide, wherein the membrane domaincomprises a transmembrane domain and an intracellular domain, andwherein the polynucleotide is an mRNA comprising a 5′ untranslatedregion (UTR), the ORF, and a 3′ UTR.

In some aspects, the disclosure provides a method of inducing ananti-tumor response in a subject in need thereof, comprisingadministering to the subject an effective amount of a lipid nanoparticlecomprising a polynucleotide comprising an open reading frame (ORF)encoding a human interleukin-12 (IL-12) polypeptide operably linked to amembrane domain, via a linker, wherein the human IL-12 polypeptidecomprises an IL-12 p40 subunit (IL-12B) polypeptide operably linked toan IL-12 p35 subunit (IL-12A) polypeptide, wherein the membrane domaincomprises a PDGFR transmembrane domain, and wherein the polynucleotideis an mRNA comprising a 5′ untranslated region (UTR), the ORF, and a 3′UTR.

In some aspects, the disclosure provides a method of inducing ananti-tumor response in a subject in need thereof, comprisingadministering to the subject an effective amount of a lipid nanoparticlecomprising a polynucleotide comprising an open reading frame (ORF)encoding a human interleukin-12 (IL-12) polypeptide operably linked to amembrane domain, via a linker, wherein the human IL-12 polypeptidecomprises an IL-12 p40 subunit (IL-12B) polypeptide operably linked toan IL-12 p35 subunit (IL-12A) polypeptide, wherein the membrane domaincomprises a CD80 transmembrane domain, and wherein the polynucleotide isan mRNA comprising a 5′ untranslated region (UTR), the ORF, and a 3′UTR.

In some aspects, the disclosure provides a method of inducing ananti-tumor response in a subject in need thereof, comprisingadministering to the subject an effective amount of a lipid nanoparticlecomprising a polynucleotide comprising an open reading frame (ORF)encoding a human interleukin-12 (IL-12) polypeptide operably linked to amembrane domain, via a linker, wherein the human IL-12 polypeptidecomprises an IL-12 p40 subunit (IL-12B) polypeptide operably linked toan IL-12 p35 subunit (IL-12A) polypeptide, wherein the membrane domaincomprises a CD8 transmembrane domain, and wherein the polynucleotide isan mRNA comprising a 5′ untranslated region (UTR), the ORF, and a 3′UTR.

In some aspects, the disclosure provides a method of inducing ananti-tumor response in a subject in need thereof, comprisingadministering to the subject an effective amount of a lipid nanoparticlecomprising a polynucleotide comprising an open reading frame (ORF)encoding a human interleukin-12 (IL-12) polypeptide operably linked to amembrane domain, via a linker, wherein the human IL-12 polypeptidecomprises an IL-12 p40 subunit (IL-12B) polypeptide operably linked toan IL-12 p35 subunit (IL-12A) polypeptide, wherein the membrane domaincomprises a CD80 transmembrane domain and a CD80 intracellular domain,and wherein the polynucleotide is an mRNA comprising a 5′ untranslatedregion (UTR), the ORF, and a 3′ UTR.

In some aspects, the disclosure provides a method of inducing ananti-tumor response in a subject in need thereof, comprisingadministering to the subject an effective amount of a lipid nanoparticlecomprising a polynucleotide comprising an open reading frame (ORF)encoding a human interleukin-12 (IL-12) polypeptide operably linked to amembrane domain, via a linker, wherein the human IL-12 polypeptidecomprises an IL-12 p40 subunit (IL-12B) polypeptide operably linked toan IL-12 p35 subunit (IL-12A) polypeptide, wherein the membrane domaincomprises a PDGFR transmembrane domain and a PDGFR intracellular domain,and wherein the polynucleotide is an mRNA comprising a 5′ untranslatedregion (UTR), the ORF, and a 3′ UTR.

In any of the foregoing methods, the polynucleotide, mRNA, composition,lipid nanoparticle lipid or pharmaceutical composition is administeredby intratumoral injection.

In any of the foregoing methods, the anti-tumor response comprises aT-cell response. In some aspects, the T-cell response comprises CD8+ Tcells.

In any of the foregoing aspects, the method further comprisesadministering to the subject an effective amount of a compositioncomprising an anti-cancer agent, or a polynucleotide comprising an ORFencoding an anti-cancer agent.

In any of the foregoing aspects, the method further comprisesadministering a second composition comprising a checkpoint inhibitorpolypeptide or polynucleotide encoding the same, and an optionalpharmaceutically acceptable carrier. In some aspects, the checkpointinhibitor polypeptide inhibits PD1, PD-L1, CTLA4, or a combinationthereof. In some aspects, the checkpoint inhibitor polypeptide is anantibody. In some aspects, the checkpoint inhibitor polypeptide is anantibody selected from an anti-CTLA4 antibody or antigen-bindingfragment thereof that specifically binds CTLA4, an anti-PD1 antibody orantigen-binding fragment thereof that specifically binds PD1, ananti-PD-L1 antibody or antigen-binding fragment thereof thatspecifically binds PD-L1, and a combination thereof. In some aspects,the checkpoint inhibitor polypeptide is an anti-PD-L1 antibody selectedfrom atezolizumab, avelumab, or durvalumab. In some aspects, thecheckpoint inhibitor polypeptide is an anti-CTLA-4 antibody selectedfrom tremelimumab or ipilimumab. In some aspects, the checkpointinhibitor polypeptide is an anti-PD1 antibody selected from nivolumab orpembrolizumab.

In any of the foregoing aspects, the composition comprising thecheckpoint inhibitor polypeptide is administered by intravenousinjection. In some aspects, the composition comprising the checkpointinhibitor polypeptide is administered once every 2 to 3 weeks.

In any of the foregoing aspects, the second composition comprising thecheckpoint inhibitor polypeptide is administered prior to, concurrentwith, or subsequent to administration of the polynucleotide, mRNA,composition, lipid nanoparticle or pharmaceutical composition.

BRIEF DESCRIPTION OF THE DRAWINGS/FIGURES

FIGS. 1A-1D show exemplary structures of tethered IL-12 polypeptideswith (FIGS. 1A and 1C) or without (FIGS. 1B and 1D) a linker between anIL-12 polypeptide (“IL12”) and a membrane domain (“MD”). The “IL12”polypeptide includes polypeptides comprising IL-12A, IL-12B, or bothIL-12A and IL-12B. “N” indicates the amino-terminus of the polypeptide,while “C” indicates the carboxy-terminus of the polypeptide.

FIGS. 2A-2E show structures of tethered murine IL-12 polypeptides,comprising an IL-12B subunit linked to an IL-12A subunit via a linker,as used in the Examples. FIG. 2A shows an IL-12 polypeptide linked to aCD8 transmembrane domain via a linker, with a V5 tag (“mIL12-8TM”). FIG.2B shows an IL-12 polypeptide linked to a PGFRB transmembrane domain viaa linker, with a V5 tag (“mIL12-PTM”). FIG. 2C shows an IL-12polypeptide linked to a CD80 transmembrane domain and intracellulardomain via a linker (“mIL12-80TID”). FIG. 2D shows an IL-12 polypeptidelinked to a CD80 transmembrane domain, with no linker, and a V5 tag(“mIL12-80TM”). FIG. 2E shows an IL-12 polypeptide linked to a CD80transmembrane and intracellular domain without a linker, and comprisinga hemagglutinin (HA) tag and IgK signal peptide (“IgK_mscIL12-80TID”).

FIG. 3 is a graph depicting in vitro expression levels of IL-12 in thesupernatant or lysates of HeLa cells after 24 hours of exposure totransfection reagent with no mRNA (“Mock”) or transfected in individualcell culture wells with the following constructs: mRNA encoding asecreted mouse IL-12 polypeptide (“mIL12AB”), mIL12-8TM, mIL12-PTM, andmIL12-80TM. The amount of IL-12 in nanograms per respective culture well(“ng/well”) is shown on the y-axis of the figure.

FIGS. 4A-4D show the in vitro bioactivity associated with varioustethered IL-12 polypeptide constructs. FIGS. 4A and 4C show the level ofproliferation of mouse splenic CD8+ T cell in relative light units(“RLU”) on the y-axis. FIGS. 4B and 4D show the amount of interferongamma (IFNγ) secretion by mouse splenic CD8+ T cells in nanograms permilliliter (“ng/ml”) on the y-axis. Proliferation of and secretion byCD8+ T cells in each of FIGS. 4A-4D was measured after 72 hours inco-culture with “Mock” transfected HeLa cells or HeLa cells transfectedin individual culture wells with mIL12AB, mIL12-8TM, or mIL12-80TM asdescribed in the brief description of FIG. 3 . Recombinant mouse IL-12(rmIL12) was also added to a subset of “Mock” cultures (“Mock+rmIL12”).Each condition in FIGS. 4A-4D represent 50,000 CD8+ T cells culturedwith a fixed number of HeLa cells from a HeLa cell culture transfectedwith one of the noted constructs and further including a fixed amount ofsupernatant from the same HeLa cell culture.

FIGS. 5A and 5B show in vitro IL-12 protein expression and bioactivityon mouse splenic CD8+ T cells of HeLa cells transfected with IL-12polypeptide constructs. FIG. 5A shows the amount of IL-12 (ng/well) inthe supernatant or lysates of “Mock” HeLa cells or HeLa cells 24 hoursafter transfection in individual cell culture wells with varioustethered murine IL-12 mRNAs. FIG. 5B shows the amount of IFNγ secretion(ng/mL) by mouse splenic CD8+ T cells after 72 hours of co-culture with“Mock” or transfected HeLa cells and supernatant.

FIGS. 6A and 6B show the plasma levels of IL-12 (FIG. 6A) and IFNγ (FIG.6B) from mice having MC38 tumors, after treatment with mIL12AB(“secreted mIL-12”) or mIL12-PTM (“tethered mIL-12”). Each graph showsplasma concentration in picograms per milliliter (“pg/mL”) over timepost dose in hours.

FIG. 7 provides graphs showing the percentage of body weight change overtime (days) post implant of MC38 tumors into mice. The vertical lineindicates the day treatment began with either negative control (NSTmRNA; left), secreted IL-12 (mIL12AB; middle) or tethered IL-12(mIL12-PTM; right).

FIGS. 8A-8E show in vivo tumor efficacy in both primary treated andsecondary untreated (i.e., distal) tumors with either a negative controlmRNA encoding an untranslatable sequence of mOX40L (“Negative Control”),or mIL12-PTM mRNA construct as described in the brief description ofFIG. 2B. FIG. 8A shows a schematic description of the MC38 dual flankmodel used in the experiments. “5e5” indicates that 5×10⁵ MC38 cellswere inoculated into the primary (right) or secondary (left) flanks toproduce the tumors. Tumors in the right flank (primary tumors) weretreated by intratumoral injection of one of the mRNAs, while tumors inthe left flank (secondary tumors (i.e., distal tumors)) did not receiveintratumoral injections. The effect of intratumoral administration ofthe mRNA in the primary tumor was determined by measuring tumor volumein both the primary tumor and the secondary tumor. The y-axes of FIGS.8B-8E show tumor volume in cubic millimeters (mm³) at the number of daysindicated on the x-axis post implant with MC38. FIG. 8B shows the effectof the negative control mRNA on the primary treated tumor. FIG. 8C showsthe effect of the negative control mRNA on the secondary tumor. FIG. 8Dshows the effect of mIL12-PTM on the primary treated tumor. FIG. 8Eshows the effect of mIL12-PTM on the secondary tumor.

FIGS. 9A-9D show structures of tethered human IL-12 polypeptides,comprising an IL-12B subunit linked to an IL-12A subunit via a linker,as used in the Examples. FIG. 9A shows an IL-12 polypeptide linked to aCD8 transmembrane domain via a linker, with a V5 tag (“hIL12-8TM”). FIG.9B shows an IL-12 polypeptide linked to a CD80 transmembrane domain andintracellular domain via a linker (“hIL12-80TID”). FIG. 9C shows anIL-12 polypeptide linked to a PGFRB transmembrane domain and truncatedintracellular domain (E570tr), via a linker (“hIL12-PTIDE570”). FIG. 9Dshows an IL-12 polypeptide linked to a PGFRB transmembrane domain andtruncated intracellular domain (G739tr), via a linker(“hIL12-PTIDG739”).

FIGS. 10A and 10B show in vitro IL-12 protein expression and bioactivityon human peripheral blood CD8+ T cells of HeLa cells transfected withIL-12 polypeptide constructs. FIG. 10A shows the amount of IL-12(ng/well) in the supernatant or lysates of “Mock” HeLa cells or HeLacells 24 hours after transfection in individual cell culture wells withvarious tethered human IL-12 mRNAs 24 hours after transfection. FIG. 10Bshows the amount of IFNγ secretion (ng/mL) by human peripheral bloodCD8+ T cells after 72 hours of co-culture with “Mock” or transfectedHeLa cells and supernatants.

FIG. 11 shows in vitro IL-12 protein expression of hIL12-80TID encodedby four different mRNA sequences. The graph shows the amount of IL-12(ng/well) in the supernatant (left) or lysates (right) of “Mock” HeLacells or HeLa cells transfected in individual wells with the variousmRNAs 24 hours after transfection.

DETAILED DESCRIPTION

The present disclosure provides a new approach to treat cancer involvingthe prevention or treatment of disease with substances (e.g., mRNAsencoding a tethered IL-12 polypeptide, which comprises an IL-12polypeptide and a membrane domain as disclosed herein) that stimulatethe immune response, i.e., immunotherapy.

In one aspect, the disclosure relates to methods of treating cancerusing a polynucleotide (e.g., mRNA) encoding a tethered IL-12polypeptide. An IL-12 polypeptide as disclosed herein comprises IL-12A,IL-12B, or both IL-12A and IL-12B. In another aspect, the disclosureprovides methods of treating cancer using a combination approach thatfeatures a polynucleotide (e.g., mRNA) encoding a tethered IL-12polypeptide and an anti-cancer agent, e.g., an immune-checkpointinhibitor, e.g., anti-PD-1 antibody, anti-PD-L1 antibody, and/oranti-CTLA-4 antibody. Without being bound by any theory, it is believedthat priming of an anti-cancer immune response is possible byadministering, e.g., intratumorally, a polynucleotide (e.g., mRNA)encoding a tethered IL-12 polypeptide in the stimulation of, forexample, T-cells and/or natural killer cells. Therefore, apolynucleotide (e.g., mRNA) encoding a tethered IL-12 polypeptide isbelieved to provide a first stimulation signal to the immune system, forexample, within the tumor environment, e.g., via intratumoral injectionof the polynucleotide (e.g., mRNA). IL-12 can also stimulate theproduction of interferon-gamma (IFN-γ) and tumor necrosis factor-alpha(TNF-α) from T cells and natural killer (NK) cells. As disclosed herein,IL-12, either directly or indirectly through IFN-γ, can also increaseexpression of PD-L1 in tumor cells, which can impair local tumorimmunity. Therefore, in some aspects, the disclosure provides a methodof treating a tumor comprising administering a polynucleotide (e.g.,mRNA) encoding a tethered IL-12 polypeptide in combination with ananti-PD-1 antibody or anti-PD-L1 antibody to block the interactionbetween PD-L1 and its receptor, i.e., PD-1. In other aspects, thedisclosure includes a method of treating a tumor comprisingadministering a polynucleotide (e.g., mRNA) encoding a tethered IL-12polypeptide in combination with an anti-CTLA-4 antibody. In furtheraspects, the disclosure provides a method of treating a tumor comprisingadministering a polynucleotide (e.g., mRNA) encoding a tethered IL-12polypeptide in combination with an anti-PD-1 antibody or anti-PD-L1antibody and an anti-CTLA-4 antibody. Some aspects of the disclosurealso include additional agents, e.g., an antibody. In other aspects, theanti-PD-1 antibody or anti-PD-L1 antibody can be administered in theform of a polynucleotide. Similarly, the anti-CTLA-4 antibody can beadministered in the form of a polynucleotide. Exemplary aspects featuretreatment with lipid nanoparticle-(LNP-) encapsulated mRNAs. Exemplaryaspects feature intratumoral administration of mRNAs in cationiclipid-based LNPs.

1. Methods of Treating Cancer

Certain aspects of the present disclosure are directed to methods ofreducing or decreasing size, mass, and/or volume of a tumor orpreventing the growth of a tumor in a subject in need thereof comprisingadministering a polynucleotide, e.g., mRNA, encoding a tethered IL-12polypeptide disclosed herein, or a vector or a host cell comprising thepolynucleotide, or a tethered IL-12 polypeptide encoded by thepolynucleotide.

In other embodiments, the present disclosure provides methods ofpromoting an anti-tumor effect (e.g., induce T cell proliferation,induce T cell infiltration in a tumor, induce a memory T cell response,increasing the number of NK cells, etc.) by administering thepolynucleotide (e.g., mRNA) encoding a tethered IL-12 polypeptide or thepolynucleotide in combination with any agents disclosed herein.

In one embodiment, the present disclosure provides a method ofactivating T cells in a subject in need thereof, inducing T cellproliferation in a subject in need thereof, inducing T cell infiltrationin a tumor of a subject in need thereof, and/or inducing a memory T cellresponse in a subject in need thereof, comprising administering to thesubject a polynucleotide encoding a tethered IL-12 polypeptide alone orin combination with a second agent, e.g., a checkpoint inhibitor, e.g.,an anti-PD-1 antibody, an anti-PD-L1 antibody, and/or an anti-CTLA-4antibody. In certain embodiments, the intratumoral administration of thepolynucleotide (e.g., mRNA) encoding a tethered IL-12 polypeptide aloneor in combination with a second agent can increase the efficacy of theanti-tumor effect (e.g., T cell infiltration in a tumor) compared toother routes of administration.

Administration of cytokines, such as IL-12, has been associated withtoxicities in treated subjects. See, e.g., Leonard, J. P. et al., Blood90: 2541-2548 (1997). In other embodiments, the present disclosureprovides a method of reducing the size of a tumor or inhibiting thegrowth of a tumor in a subject in need thereof with reduced toxicity,comprising administering to the subject a polynucleotide (e.g., mRNA)encoding a tethered IL-12 polypeptide disclosed herein. In someembodiments, the administration exhibits reduced toxicity compared to anadministration of a reference polynucleotide (e.g., mRNA) encoding anIL-12 polypeptide that is not tethered. In some embodiments, the reducedtoxicity is a reduction in a toxicity or a toxic effect selected fromthe group consisting of: systemic toxicity, sepsis-like syndrome, septicshock, cachexia, loss of weight, muscle atrophy, fatigue, weakness,significant loss of appetite, hepatotoxicity, a decrease in circulatingleukocytes, thrombocytopenia, anemia, dyspnea, stomatitis, leukopenia,hyperbilirubinemia, elevations in transaminases, thrombocytopenia, organfailure, respiratory failure, liver failure, renal failure,gastrointestinal bleeding, and combinations thereof.

In one embodiment, activated T cells in the subject reduce the size of atumor or inhibit the growth of a tumor in the subject. Activation of Tcells can be measured using applications in the art such as measuring Tcell proliferation; measuring cytokine production with enzyme-linkedimmunosorbent assays (ELISA) or enzyme-linked immunospot assays(ELISPOT); or detection of cell-surface markers associated with T cellactivation (e.g., CD69, CD40L, CD137, CD25, CD71, CD26, CD27, CD28,CD30, CD154, and CD134) with techniques such as flow cytometry.

In one embodiment, T cell proliferation in the subject is directed to ananti-tumor immune response in the subject. In another aspect, the T cellproliferation in the subject reduces or decreases the size of a tumor orinhibits the growth of a tumor in the subject. T cell proliferation canbe measured using applications in the art such as cell counting,viability staining, optical density assays, or detection of cell-surfacemarkers associated with T cell activation (e.g., CD69, CD40L, CD137,CD25, CD71, CD26, CD27, CD28, CD30, CD154, and CD134) with techniquessuch as flow cytometry.

In one embodiment, T cell infiltration in a tumor of the subject isdirected to an anti-tumor immune response in the subject. In anotheraspect, the T cell infiltration in a tumor of the subject reduces ordecreases the size of a tumor or inhibits the growth of a tumor in thesubject. T cell infiltration in a tumor can be measured usingapplications in the art such as tissue sectioning and staining for cellmarkers, measuring local cytokine production at the tumor site, ordetection of T cell-surface markers with techniques such as flowcytometry.

In one embodiment, the memory T cell response in the subject is directedto an anti-tumor immune response in the subject. In another aspect, thememory T cell response in the subject reduces or decreases the size of atumor or inhibits the growth of a tumor in the subject. A memory T cellresponse can be measured using applications in the art such as measuringT cell markers associated with memory T cells, measuring local cytokineproduction related to memory immune response, or detecting memory Tcell-surface markers with techniques such as flow cytometry.

In certain embodiments, the T cells activated by the present methods areCD4⁺ cells, CD8⁺ cells, CD62⁺ (L-selectin⁺) cells, CD69⁺ cells, CD40L⁺cells, CD137⁺ cells, CD25⁺ cells, CD71⁺ cells, CD26⁺ cells, CD27⁺ cells,CD28⁺ cells, CD30⁺ cells, CD45⁺ cells, CD45RA⁺ cells, CD45RO⁺ cells,CD11b⁺ cells, CD154⁺ cells, CD134⁺ cells, CXCR3⁺ cells, CCR4⁺ cells,CCR6⁺ cells, CCR7⁺ cells, CXCR5⁺ cells, Crth2⁺ cells, gamma delta Tcells, or any combination thereof. In some embodiments, the T cellsactivated by the present methods are Thi cells. In other embodiments,the T cells activated by the present methods are Th₂ cells. In otherembodiments, the T cells activated by the present methods are cytotoxicT cells.

In some embodiments, the infiltrating T cells induced by the presentmethods are CD4⁺ cells, CD8⁺ cells, CD62⁺ (L-selectin⁺) cells, CD69⁺cells, CD40L⁺ cells, CD137⁺ cells, CD25⁺ cells, CD71⁺ cells, CD26⁺cells, CD27⁺ cells, CD28⁺ cells, CD30⁺ cells, CD45⁺ cells, CD45RA⁺cells, CD45RO⁺ cells, CD11b⁺ cells, CD154⁺ cells, CD134⁺ cells, CXCR3⁺cells, CCR4⁺ cells, CCR6⁺ cells, CCR7⁺ cells, CXCR5⁺ cells, Crth2⁺cells, gamma delta T cells, or any combination thereof. In someembodiments, the infiltrating T cells induced by the present methods areThi cells. In other embodiments, the infiltrating T cells induced by thepresent methods are Th₂ cells. In other embodiments, the infiltrating Tcells induced by the present methods are cytotoxic T cells.

In some embodiments, the memory T cells induced by the present methodsare CD4⁺ cells, CD8⁺ cells, CD62⁺ (L-selectin⁺) cells, CD69⁺ cells,CD40L⁺ cells, CD137⁺ cells, CD25⁺ cells, CD71⁺ cells, CD26⁺ cells, CD27⁺cells, CD28⁺ cells, CD30⁺ cells, CD45⁺ cells, CD45RA⁺ cells, CD45RO⁺cells, CD11b⁺ cells, CD154⁺ cells, CD134⁺ cells, CXCR3⁺ cells, CCR4⁺cells, CCR6⁺ cells, CCR7⁺ cells, CXCR5⁺ cells, Crth2⁺ cells, gamma deltaT cells, or any combination thereof. In some embodiments, the memory Tcells induced by the present methods are Thi cells. In otherembodiments, the memory T cells induced by the present methods are Th₂cells. In other embodiments, the memory T cells induced by the presentmethods are cytotoxic T cells.

In certain embodiments, the disclosure provides a method of inducing anadaptive immune response, an innate immune response, or both adaptiveand innate immune response against a tumor, comprising administering apolynucleotide, e.g., mRNA, encoding a tethered IL-12 polypeptide aloneor in combination with a second agent, e.g., a checkpoint inhibitor,e.g., an anti-PD-1 antibody, an anti-PD-L1 antibody, and/or ananti-CTLA-4 antibody, and/or any other agents disclosed herein. In someembodiments, the disclosure provides a method of inducing an adaptiveimmune response, an innate immune response, or both adaptive and innateimmune response against a tumor, comprising administering apolynucleotide, e.g., mRNA, encoding a tethered IL-12 polypeptide incombination with a second agent, e.g., a checkpoint inhibitor, e.g., ananti-PD-1 antibody, an anti-PD-L1 antibody, and/or an anti-CTLA-4antibody, and/or any other agents disclosed herein. In some embodiments,the checkpoint inhibitor can be a polynucleotide (e.g., mRNA) encodingan antibody or an antigen-binding portion thereof, e.g., an anti-PD-1antibody, an anti-PD-L1 antibody, and/or an anti-CTLA-4 antibody. Insome embodiments, the disclosure provides a method of inducing anadaptive immune response, an innate immune response, or both adaptiveand innate immune response against a tumor, comprising administering apolynucleotide, e.g., mRNA, encoding a tethered IL-12 polypeptide.

The present disclosure further provides a method of increasing thenumber of Natural Killer (NK) cells in a subject in need thereof,comprising administering a polynucleotide comprising an mRNA encoding atethered IL-12 polypeptide alone or in combination with a second agent,e.g., a checkpoint inhibitor, e.g., an anti-PD-1 antibody, an anti-PD-L1antibody, and/or an anti-CTLA-4 antibody, and/or any other agentsdisclosed herein. In some embodiments, the disclosure provides a methodof increasing the number of Natural Killer (NK) cells in a subject inneed thereof, comprising administering a polynucleotide comprising anmRNA encoding a tethered IL-12 polypeptide. In some embodiments, thedisclosure provides a method of increasing the number of Natural Killer(NK) cells in a subject in need thereof, comprising administering apolynucleotide comprising an mRNA encoding a tethered IL-12 polypeptidein combination with a second agent, e.g., a checkpoint inhibitor, e.g.,an anti-PD-1 antibody, an anti-PD-L1 antibody, and/or an anti-CTLA-4antibody, and/or any other agents disclosed herein. In one aspect, theincrease in the number of NK cells in the subject is directed to ananti-tumor immune response in the subject. In another aspect, theincrease in the number of NK cells in the subject reduces or decreasesthe size of a tumor or inhibits the growth of a tumor in the subject.Increases in the number of NK cells in a subject can be measured usingapplications in the art such as detection of NK cell-surface markers(e.g., CD335/NKp46; CD336/NKp44; CD337/NPp30) or intracellular NK cellmarkers (e.g., perforin; granzymes; granulysin).

In certain embodiments, the present disclosure is also directed to amethod of increasing IFNγ expression in a subject having tumorcomprising administering a polynucleotide encoding a tethered IL-12polypeptide alone or in combination with a second agent, e.g., acheckpoint inhibitor, e.g., an anti-PD-1 antibody, an anti-PD-L1antibody, and/or an anti-CTLA-4 antibody, and/or any other agentsdisclosed herein. In some embodiments, the disclosure provides a methodof increasing IFNγ expression in a subject having tumor comprisingadministering a polynucleotide encoding a tethered IL-12 polypeptide. Insome embodiments, the disclosure provides a method of increasing IFNγexpression in a subject having tumor comprising administering apolynucleotide encoding a tethered IL-12 polypeptide in combination witha second agent, e.g., a checkpoint inhibitor, e.g., an anti-PD-1antibody, an anti-PD-L1 antibody, and/or an anti-CTLA-4 antibody, and/orany other agents disclosed herein.

Other embodiments also include a method of increasing expression ofIFNγ, TNFα, IL-10, IL-13, IL-15/15R, IL-27, MIP-1β, MIP-1α, MCP-1,MCP-3, M-CSF, IL-4, IL-5, or any combination thereof in a subject havingtumor comprising administering a polynucleotide, e.g., mRNA, encoding atethered IL-12 polypeptide alone or in combination with another agentdisclosed herein. In yet other embodiments, the methods of the presentdisclosure can include methods of inducing expression of GM-CSF, IL-18,IL-3, RANTES, IL-6, or any combination thereof.

The polynucleotide encoding a tethered IL-12 polypeptide can beformulated as a pharmaceutical composition that is suitable foradministration either directly or indirectly to tumors. The term “tumor”is used herein in a broad sense and refers to any abnormal new growth oftissue that possesses no physiological function and arises fromuncontrolled usually rapid cellular proliferation. The term “tumor” asused herein relates to both benign tumors and to malignant tumors.

Certain aspects of the disclosure provide methods of intratumorallyadministering a single dose of a polynucleotide, e.g., mRNA, encoding atethered IL-12 polypeptide alone or in combination with any agentsdisclosed herein. In some embodiments, the disclosure provides methodsof intratumorally administering a single dose of a polynucleotide, e.g.,mRNA, encoding a tethered IL-12 polypeptide. In some embodiments, thedisclosure provides methods of intratumorally administering a singledose of a polynucleotide, e.g., mRNA, encoding a tethered IL-12polypeptide alone or in combination with any agents disclosed herein. Insuch embodiments, an mRNA encoding a tethered IL-12 polypeptide can beadministered only once while the other agent can be administeredregularly, following its regular dosing schedule. In certainembodiments, a checkpoint inhibitor, e.g., an anti-PD-1 antibody, ananti-PD-L1 antibody, and/or an anti-CTLA-4 antibody, is administeredprior to administration of a polynucleotide, e.g., mRNA, encoding atethered IL-12 polypeptide. In some embodiments, the polynucleotide isformulated in a lipid nanoparticle, e.g., Compound 18 based lipidnanoparticle, disclosed herein. Not being bound by any theory, in someaspects, the intratumoral delivery of a polynucleotide encoding atethered IL-12 polypeptide and/or the lipid nanoparticle formulationdisclosed herein allows single dose administration that is sufficientfor the dose to trigger anti-tumor efficacy and treat the tumor. Giventhe potential toxicity of IFNγ induced by IL-12, this single dosingregimen of the disclosed polynucleotide can be beneficial to thesubjects in need of the treatment.

In certain embodiments, the method comprises administering a single doseof a polynucleotide, e.g., mRNA, encoding a tethered IL-12 polypeptidein combination with a second agent, e.g., an anti-PD-1 antibody, ananti-PD-L1 antibody, and/or an anti-CTLA-4 antibody, which can be givenalso in single administration or multiple administrations following itsregular (e.g., approved) schedule. In other embodiments, the methodcomprises not more than two administrations of a polynucleotide, e.g.,mRNA, encoding a tethered IL-12 polypeptide, not more than threeadministrations of a polynucleotide, e.g., mRNA, encoding a tetheredIL-12 polypeptide, not more than four administrations of apolynucleotide, e.g., mRNA, encoding a tethered IL-12 polypeptide, ornot more than five administrations of a polynucleotide, e.g., mRNA,encoding a tethered IL-12 polypeptide, optionally in combination with acheckpoint inhibitor, e.g., an anti-PD-1 antibody, an anti-PD-L1antibody, and/or an anti-CTLA-4 antibody.

In other embodiments, the present methods can result in abscopaleffects, e.g., a treatment of tumor where localized treatment of atumor, e.g., intratumoral delivery, by a polynucleotide, e.g., mRNA,encoding a tethered IL-12 polypeptide causes not only a shrinking of thetreated tumor, but also a shrinking of tumors outside the scope of thelocalized treatment (“distal tumor”).

In some embodiments, the administering of a polynucleotide encoding atethered IL-12 polypeptide (alone or in combination with an anti-PD-L1antibody, an anti-PD-1 antibody, or an anti-CTLA-4 antibody) increasesan effector to suppressor T cell ratio in the tumor. In certainembodiments, the effector to suppressor T cell ratio is characterized bythe ratio of (i) CD8+, CD4+, or CD8+/CD4+ T cells to (ii) Treg cells ina subject. In certain embodiments, the increase in the effector tosuppressor T cell ratio correlates with an increase in the number ofCD8+ T cells. In some embodiments, the increase in the effector tosuppressor T cell ratio correlates with an increase in the number ofCD4+ T cells. In some embodiments, the increase in the effector tosuppressor T cell ratio correlates with an increase in the number ofCD8+/CD4+ T cells. In some embodiments, the increase in the effector tosuppressor T cell ratio correlates with a decrease in the number of Tregcells.

In some embodiments, the effector to suppressor T cell ratio, e.g., theCD8⁺ T cell to Treg cell ratio, following administration of apolynucleotide encoding a tethered IL-12 polypeptide (alone or incombination with an anti-PD-L1 antibody, an anti-PD-1 antibody, and/oran anti-CTLA-4 antibody or a polynucleotide encoding the same) is atleast about 1.5:1, at least about 2:1, at least about 2.5:1, at leastabout 3:1, at least about 3.5:1, at least about 3.5:1, at least about4:1, at least about 4.5:1, at least about 5:1, at least about 6:1, atleast about 7:1, at least about 8:1, at least about 9:1, at least about10:1, at least about 15:1, at least about 20:1, at least about 25:1, atleast about 30:1, at least about 35:1, at least about 40:1, at leastabout 45:1, at least about 50:1, at least about 60:1, at least about70:1, at least about 80:1, at least about 90:1, at least about 100:1, atleast about 110:1, at least about 120:1, at least about 130:1, at leastabout 140:1, at least about 150:1, at least about 200:1, at least about250:1, or at least about 500:1.

In some embodiments, the effector to suppressor T cell ratio, e.g., theCD8⁺ T cell to Treg cell ratio, following administration of apolynucleotide encoding a tethered IL-12 polypeptide (alone or incombination with an anti-PD-L1 antibody, an anti-PD-1 antibody, and/oran anti-CTLA-4 antibody or a polynucleotide encoding the same) is atleast about 1.5, at least about 2, at least about 2.5, at least about 3,at least about 3.5, at least about 3.5, at least about 4, at least about4.5, at least about 5, at least about 6, at least about 7, at leastabout 8, at least about 9, at least about 10, at least about 15, atleast about 20, at least about 25, at least about 30, at least about 35,at least about 40, at least about 45, at least about 50, at least about60, at least about 70, at least about 80, at least about 90, at leastabout 100, at least about 110, at least about 120, at least about 130,at least about 140, at least about 150, at least about 200, at leastabout 250, or at least about 500.

In one embodiment, the increase in the effector to suppressor T cellratio in the tumor is directed to an anti-tumor immune response in thesubject. In another aspect, the increase in the effector to suppressor Tcell ratio in the tumor reduces or decreases the size of a tumor orinhibits the growth of a tumor in the subject. The effector tosuppressor T cell ratio in the tumor can be measured using applicationsin the art such as measuring the ratio of CD8+, CD4+, or CD8+/CD4+ Tcells to Treg cells, using any methods known in the art including IHCand/or flow cytometry.

The delivery of the polynucleotide encoding a tethered IL-12 polypeptideto a tumor using a pharmaceutical composition for intratumoraladministration disclosed herein can:

-   -   (a) increase the retention of the polynucleotide in the tumor;    -   (b) increase the levels of expressed polypeptide in the tumor        compared to the levels of expressed polypeptide in peritumoral        tissue;    -   (c) decrease leakage of the polynucleotide or expressed product        to off-target tissue (e.g., peritumoral tissue, or to distant        locations, e.g., liver tissue); or,    -   (d) any combination thereof,    -   wherein the increase or decrease observed for a certain property        is relative to a corresponding reference composition (e.g.,        composition in which compounds of formula (I) are not present or        have been substituted by another ionizable amino lipid, e.g.,        MC3).

In one embodiment, a decrease in leakage can be quantified as decreasein the ratio of polypeptide expression in the tumor to polypeptideexpression in non-tumor tissues, such as peritumoral tissue or toanother tissue or organ, e.g., liver tissue.

Delivery of a polynucleotide, e.g., mRNA, encoding a tethered IL-12polypeptide to a tumor involves administering a pharmaceuticalcomposition disclosed herein, e.g., in nanoparticle form, including thepolynucleotide, e.g., mRNA, encoding a tethered IL-12 polypeptide to asubject, where administration of the pharmaceutical composition involvescontacting the tumor with the composition.

In the instance that the polynucleotide encoding a tethered IL-12polypeptide is an mRNA, upon contacting a cell in the tumor with thepharmaceutical composition, a translatable mRNA may be translated in thecell to produce a polypeptide of interest. However, mRNAs that aresubstantially not translatable may also be delivered to tumors.Substantially non-translatable mRNAs may be useful as vaccines and/ormay sequester translational components of a cell to reduce expression ofother species in the cell.

The pharmaceutical compositions disclosed herein can increase specificdelivery. As used herein, the term “specific delivery,” means deliveryof more (e.g., at least 1.5 fold more, at least 2-fold more, at least3-fold more, at least 4-fold more, at least 5-fold more, at least 6-foldmore, at least 7-fold more, at least 8-fold more, at least 9-fold more,or at least 10-fold more) of a polynucleotide, e.g., mRNA, encoding atethered IL-12 polypeptide by pharmaceutical composition disclosedherein (e.g., in nanoparticle form) to a target tissue of interest(e.g., a tumor) compared to an off-target tissue (e.g., mammalianliver).

The level of delivery of a nanoparticle to a particular tissue may bemeasured, for example, by comparing

-   -   (i) the amount of protein expressed from a polynucleotide, e.g.,        mRNA, encoding a tethered IL-12 polypeptide in a tissue to the        weight of said tissue;    -   (ii) comparing the amount of the polynucleotide, e.g., mRNA, in        a tissue to the weight of said tissue; or    -   (iii) comparing the amount of protein expressed from a        polynucleotide, e.g., mRNA, encoding a tethered IL-12        polypeptide in a tissue to the amount of total protein in said        tissue.

Specific delivery to a tumor or a particular class of cells in the tumorimplies that a higher proportion of pharmaceutical composition includinga polynucleotide, e.g., mRNA, encoding a tethered IL-12 polypeptide isdelivered to the target destination (e.g., target tissue) relative toother off-target destinations upon administration of a pharmaceuticalcomposition to a subject.

The present disclosure also provides methods to deliver intratumorally apolynucleotide, e.g., mRNA, encoding a tethered IL-12 polypeptide when apharmaceutical composition comprising the polynucleotides disclosedherein (e.g., in nanoparticle form) are administered to a tumor. Theintratumoral administration can show one or more properties selectedfrom:

-   -   (i) increased retention of the polynucleotide, e.g., mRNA,        encoding a tethered IL-12 polypeptide in the tumor;    -   (ii) increased levels of expressed polypeptide in the tumor        compared to the levels of expressed polypeptide in peritumoral        tissue;    -   (iii) decreased leakage of the polynucleotide, e.g., mRNA, or        expressed product to off-target tissue (e.g., peritumoral        tissue, or to distant locations, e.g., liver tissue); and,    -   (iv) any combination thereof, wherein the increase or decrease        observed for a certain property is relative to a corresponding        reference composition (e.g., composition in which compounds of        formula (I) are not present or have been substituted by another        ionizable amino lipid, e.g., MC3).    -   In one embodiment, a decrease in leakage can be quantified as        decrease in the ratio of polypeptide expression in the tumor to        polypeptide expression in non-tumor tissues, such as peritumoral        tissue or to another tissue or organ, e.g., liver tissue.

In some embodiments, another improvement in delivery caused as a resultof using the pharmaceutical compositions disclosed herein is a reductionin immune response with respect to the immune response observed whenother lipid components are used to deliver the same a therapeutic agentor polynucleotide encoding a therapeutic agent.

Accordingly, the present disclosure provides a method of increasingretention of a therapeutic agent (e.g., a polypeptide administered aspart of the pharmaceutical composition) in a tumor tissue in a subject,comprising administering intratumorally to the tumor tissue apharmaceutical composition disclosed herein, wherein the retention ofthe therapeutic agent in the tumor tissue is increased compared to theretention of the therapeutic agent in the tumor tissue afteradministering a corresponding reference composition.

Also provided is a method of increasing retention of a polynucleotide ina tumor tissue in a subject, comprising administering intratumorally tothe tumor tissue a pharmaceutical composition disclosed herein, whereinthe retention of the polynucleotide in the tumor tissue is increasedcompared to the retention of the polynucleotide in the tumor tissueafter administering a corresponding reference composition.

Also provided is a method of increasing retention of an expressedpolypeptide in a tumor tissue in a subject, comprising administering tothe tumor tissue a pharmaceutical composition disclosed herein, whereinthe pharmaceutical composition comprises a polynucleotide encoding theexpressed polypeptide, and wherein the retention of the expressedpolypeptide in the tumor tissue is increased compared to the retentionof the polypeptide in the tumor tissue after administering acorresponding reference composition.

The present disclosure also provides a method of decreasing expressionleakage of a polynucleotide administered intratumorally to a subject inneed thereof, comprising administering the polynucleotide intratumorallyto the tumor tissue as a pharmaceutical composition disclosed herein,wherein the expression level of the polypeptide in non-tumor tissue isdecreased compared to the expression level of the polypeptide innon-tumor tissue after administering a corresponding referencecomposition.

Also provided is a method of decreasing expression leakage of atherapeutic agent (e.g., a polypeptide administered as part of thepharmaceutical composition) administered intratumorally to a subject inneed thereof, comprising administering the therapeutic agentintratumorally to the tumor tissue as a pharmaceutical compositiondisclosed herein, wherein the amount of therapeutic agent in non-tumortissue is decreased compared to the amount of therapeutic in non-tumortissue after administering a corresponding reference composition.

Also provided is a method of decreasing expression leakage of anexpressed polypeptide in a tumor in a subject, comprising administeringto the tumor tissue a pharmaceutical composition disclosed herein,wherein the pharmaceutical composition comprises a polynucleotideencoding the expressed polypeptide, and wherein the amount of expressedpolypeptide in non-tumor tissue is decreased compared to the amount ofexpressed polypeptide in non-tumor tissue after administering acorresponding reference composition.

In some embodiments, the non-tumoral tissue is peritumoral tissue. Inother embodiments, the non-tumoral tissue is liver tissue.

The present disclosure also provides a method to reduce or prevent theimmune response caused by the intratumoral administration of apharmaceutical composition, e.g., a pharmaceutical compositioncomprising lipids known in the art, by replacing one or all the lipidsin such composition with a compound of Formula (I). For example, theimmune response caused by the administration of a polynucleotide, e.g.,mRNA, encoding a tethered IL-12 polypeptide in a pharmaceuticalcomposition comprising MC3 (or other lipids known in the art) can beprevented (avoided) or ameliorated by replacing MC3 with a compound ofFormula (I), e.g., Compound 18.

In some embodiments, the immune response observed after apolynucleotide, e.g., mRNA, encoding a tethered IL-12 polypeptide isadministered in a pharmaceutical composition disclosed herein is notelevated compared to the immune response observed when the therapeuticagent or a polynucleotide, e.g., mRNA, encoding a tethered IL-12polypeptide is administered in phosphate buffered saline (PBS) oranother physiological buffer solution (e.g., Ringer's solution, Tyrode'ssolution, Hank's balanced salt solution, etc.).

In some embodiments, the immune response observed after a therapeuticagent or a polynucleotide, e.g., mRNA, encoding a tethered IL-12polypeptide is administered in a pharmaceutical composition disclosedherein is not elevated compared to the immune response observed when PBSor another physiological buffer solution is administered alone.

In some embodiments, no immune response is observed when apharmaceutical composition disclosed herein is administeredintratumorally to a subject.

Accordingly, the present disclosure also provides a method of deliveringa therapeutic agent or a polynucleotide, e.g., mRNA, encoding a tetheredIL-12 polypeptide to a subject in need thereof, comprising administeringintratumorally to the subject a pharmaceutical composition disclosedherein, wherein the immune response caused by the administration of thepharmaceutical composition is not elevated compared to the immuneresponse caused by the intratumoral administration of

-   -   (i) PBS alone, or another physiological buffer solution (e.g.,        Ringer's solution, Tyrode's solution, Hank's balanced salt        solution, etc.);    -   (ii) the therapeutic agent or polynucleotide, e.g., mRNA,        encoding a tethered IL-12 polypeptide in PBS or another        physiological buffer solution; or,    -   (iii) a corresponding reference composition, i.e., the same        pharmaceutical composition in which the compound of Formula (I)        is substituted by another ionizable amino lipid, e.g., MC3.

In certain embodiments, the administration treats a cancer.

The polynucleotide (e.g., mRNA) of the present disclosure can beadministered in any route available, including, but not limited to,intratumoral, enteral, gastroenteral, epidural, oral, transdermal,epidural (peridural), intracerebral (into the cerebrum),intracerebroventricular (into the cerebral ventricles), epicutaneous(application onto the skin), intradermal, (into the skin itself),subcutaneous (under the skin), nasal administration (through the nose),intravenous (into a vein), intraperitoneal (into the peritoneum),intraarterial (into an artery), intramuscular (into a muscle),intracardiac (into the heart), intraosseous infusion (into the bonemarrow), intrathecal (into the spinal canal), intraperitoneal, (infusionor injection into the peritoneum), intravesical infusion, intravitreal,(through the eye), intracavernous injection, (into the base of thepenis), intravaginal administration, intrauterine, extra-amnioticadministration, transdermal (diffusion through the intact skin forsystemic distribution), transmucosal (diffusion through a mucousmembrane), insufflation (snorting), sublingual, sublabial, enema, eyedrops (onto the conjunctiva), or in ear drops. In other embodiments, thepolynucleotide, e.g., mRNA, of the present disclosure is administeredparenterally (e.g., includes subcutaneous, intravenous, intraperitoneal,intratumoral, intramuscular, intra-articular, intra-synovial,intrasternal, intrathecal, intrahepatic, intralesional and intracranialinjection or infusion techniques), intraventricularly, orally, byinhalation spray, topically, rectally, nasally, buccally, vaginally orvia an implanted reservoir. In particular embodiments, thepolynucleotide, composition, or polypeptide is administeredsubcutaneously, intravenously, intraperitoneally, intratumorally,intramuscularly, intra-articularly, intra-synovially, intrasternally,intrathecally, intrahepatically, intradermally, intralesionally,intracranially, intraventricularly, orally, by inhalation spray,topically, rectally, nasally, buccally, vaginally or via an implantedreservoir. In one particular embodiment, the polynucleotide (e.g., mRNA)of the present disclosure is administered intratumorally.

In some embodiments, the polynucleotide, e.g., mRNA, is delivered by adevice comprising a pump, patch, drug reservoir, short needle device,single needle device, multiple needle device, micro-needle device, jetinjection device, ballistic powder/particle delivery device, catheter,lumen, cryoprobe, cannula, microcanular, or devices utilizing heat, RFenergy, electric current, or any combination thereof.

Other aspects of the present disclosure relate to transplantation ofcells containing polynucleotides to a mammalian subject. Administrationof cells to mammalian subjects is known to those of ordinary skill inthe art, and includes, but is not limited to, local implantation (e.g.,topical or subcutaneous administration), organ delivery or systemicinjection (e.g., intravenous injection or inhalation), and theformulation of cells in pharmaceutically acceptable carriers.

In some embodiments, the polynucleotide, e.g., mRNA, is administered atan amount between about 0.10 μg per tumor and about 1000 mg per tumor.

In some embodiments, the administration of the polynucleotide, e.g.,mRNA, pharmaceutical composition or formulation of the disclosureresults in expression of IL-12 in cells of the subject. In someembodiments, administering the polynucleotide, e.g., mRNA,pharmaceutical composition or formulation of the disclosure results inan increase of IL-12 activity in the subject. For example, in someembodiments, the polynucleotides of the present disclosure are used inmethods of administering a composition or formulation comprising an mRNAencoding a tethered IL-12 polypeptide to a subject, wherein the methodresults in an increase of IL-12 activity in at least some cells of asubject.

In some embodiments, the administration of a composition or formulationcomprising an mRNA encoding a tethered IL-12 polypeptide to a subjectresults in an increase of IL-12 activity in cells subject to a level atleast 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 55%, atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, or to 100% or more of theactivity level expected in a normal subject.

Other embodiments of the disclosure also provide a method of treating acancer in a subject in need thereof comprising administering (e.g.,intratumorally, intraperitoneally, or intravenously) a polynucleotidecomprising an mRNA encoding a tethered IL-12 polypeptide with one ormore anti-cancer agents to the subject.

In some embodiments, the polynucleotides (e.g., mRNA) encoding atethered IL-12 polypeptide of the present disclosure can be used toreduce or decrease the size of a tumor or inhibit growth of a tumor in asubject in need thereof.

In some embodiments, the tumor is associated with a disease, disorder,and/or condition. In a particular embodiment, the disease, disorder,and/or condition is a cancer. Thus, in one aspect, the administration ofthe polynucleotide (e.g., mRNA) encoding a tethered IL-12 polypeptidetreats a cancer.

A “cancer” refers to a broad group of various diseases characterized bythe uncontrolled growth of abnormal cells in the body. Unregulated celldivision and growth results in the formation of malignant tumors thatinvade neighboring tissues and can also metastasize to distant parts ofthe body through the lymphatic system or bloodstream. A “cancer” or“cancer tissue” can include a tumor at various stages. In certainembodiments, the cancer or tumor is stage 0, such that, e.g., the canceror tumor is very early in development and has not metastasized. In someembodiments, the cancer or tumor is stage I, such that, e.g., the canceror tumor is relatively small in size, has not spread into nearby tissue,and has not metastasized. In other embodiments, the cancer or tumor isstage II or stage III, such that, e.g., the cancer or tumor is largerthan in stage 0 or stage I, and it has grown into neighboring tissuesbut it has not metastasized, except potentially to the lymph nodes. Inother embodiments, the cancer or tumor is stage IV, such that, e.g., thecancer or tumor has metastasized. Stage IV can also be referred to asadvanced or metastatic cancer.

In some aspects, the cancer can include, but is not limited to, adrenalcortical cancer, advanced cancer, anal cancer, aplastic anemia, bileduct cancer, bladder cancer, bone cancer, bone metastasis, brain tumors,brain cancer, breast cancer, childhood cancer, cancer of unknown primaryorigin, Castleman disease, cervical cancer, colon/rectal cancer,endometrial cancer, esophagus cancer, Ewing family of tumors, eyecancer, gallbladder cancer, gastrointestinal carcinoid tumors,gastrointestinal stromal tumors, gestational trophoblastic disease,Hodgkin disease, Kaposi sarcoma, renal cell carcinoma, laryngeal andhypopharyngeal cancer, acute lymphocytic leukemia, acute myeloidleukemia, chronic lymphocytic leukemia, chronic myeloid leukemia,chronic myelomonocytic leukemia, liver cancer, hepatocellular carcinoma(HCC), non-small cell lung cancer, small cell lung cancer, lungcarcinoid tumor, lymphoma of the skin, malignant mesothelioma, multiplemyeloma, myelodysplastic syndrome, nasal cavity and paranasal sinuscancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, oralcavity and oropharyngeal cancer, osteosarcoma, ovarian cancer,pancreatic cancer, penile cancer, pituitary tumors, prostate cancer,retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma inadult soft tissue, basal and squamous cell skin cancer, melanoma, smallintestine cancer, stomach cancer, testicular cancer, throat cancer,thymus cancer, thyroid cancer, uterine sarcoma, vaginal cancer, vulvarcancer, Waldenstrom macroglobulinemia, Wilms tumor, secondary cancerscaused by cancer treatment, and any combination thereof.

In some aspects, the tumor is a solid tumor. A “solid tumor” includes,but is not limited to, sarcoma, melanoma, carcinoma, or other solidtumor cancer. “Sarcoma” refers to a tumor which is made up of asubstance like the embryonic connective tissue and is generally composedof closely packed cells embedded in a fibrillar or homogeneoussubstance. Sarcomas include, but are not limited to, chondrosarcoma,fibrosarcoma, lymphosarcoma, melanosarcoma, myxosarcoma, osteosarcoma,Abemethy's sarcoma, adipose sarcoma, liposarcoma, alveolar soft partsarcoma, ameloblastic sarcoma, botryoid sarcoma, chloroma sarcoma,chorio carcinoma, embryonal sarcoma, Wilms' tumor sarcoma, endometrialsarcoma, stromal sarcoma, Ewing's sarcoma, fascial sarcoma, fibroblasticsarcoma, giant cell sarcoma, granulocytic sarcoma, Hodgkin's sarcoma,idiopathic multiple pigmented hemorrhagic sarcoma, immunoblastic sarcomaof B cells, lymphoma, immunoblastic sarcoma of T-cells, Jensen'ssarcoma, Kaposi's sarcoma, Kupffer cell sarcoma, angiosarcoma,leukosarcoma, malignant mesenchymoma sarcoma, parosteal sarcoma,reticulocytic sarcoma, Rous sarcoma, serocystic sarcoma, synovialsarcoma, or telangiectaltic sarcoma.

The term “melanoma” refers to a tumor arising from the melanocyticsystem of the skin and other organs. Melanomas include, for example,acra-lentiginous melanoma, amelanotic melanoma, benign juvenilemelanoma, Cloudman's melanoma, S91 melanoma, Harding-Passey melanoma,juvenile melanoma, lentigo maligna melanoma, malignant melanoma,metastatic melanoma, nodular melanoma, subungal melanoma, or superficialspreading melanoma.

The term “carcinoma” refers to a malignant new growth made up ofepithelial cells tending to infiltrate the surrounding tissues and giverise to metastases. Exemplary carcinomas include, e.g., acinarcarcinoma, acinous carcinoma, adenocystic carcinoma, adenoid cysticcarcinoma, carcinoma adenomatosum, carcinoma of adrenal cortex, alveolarcarcinoma, alveolar cell carcinoma, basal cell carcinoma, carcinomabasocellulare, basaloid carcinoma, basosquamous cell carcinoma,bronchioalveolar carcinoma, bronchiolar carcinoma, bronchogeniccarcinoma, cerebriform carcinoma, cholangiocellular carcinoma, chorioniccarcinoma, colloid carcinoma, comedo carcinoma, corpus carcinoma,cribriform carcinoma, carcinoma en cuirasse, carcinoma cutaneum,cylindrical carcinoma, cylindrical cell carcinoma, duct carcinoma,carcinoma durum, embryonal carcinoma, encephaloid carcinoma, epiermoidcarcinoma, carcinoma epitheliale adenoides, exophytic carcinoma,carcinoma ex ulcere, carcinoma fibrosum, gelatiniform carcinoma,gelatinous carcinoma, giant cell carcinoma, carcinoma gigantocellulare,glandular carcinoma, granulosa cell carcinoma, hair-matrix carcinoma,hematoid carcinoma, hepatocellular carcinoma, Hurthle cell carcinoma,hyaline carcinoma, hypemephroid carcinoma, infantile embryonalcarcinoma, carcinoma in situ, intraepidermal carcinoma, intraepithelialcarcinoma, Krompecher's carcinoma, Kulchitzky-cell carcinoma, large-cellcarcinoma, lenticular carcinoma, carcinoma lenticulare, lipomatouscarcinoma, lymphoepithelial carcinoma, carcinoma medullare, medullarycarcinoma, melanotic carcinoma, carcinoma molle, mucinous carcinoma,carcinoma muciparum, carcinoma mucocellulare, mucoepidernoid carcinoma,carcinoma mucosum, mucous carcinoma, carcinoma myxomatodes,naspharyngeal carcinoma, oat cell carcinoma, carcinoma ossificans,osteoid carcinoma, papillary carcinoma, periportal carcinoma,preinvasive carcinoma, prickle cell carcinoma, pultaceous carcinoma,renal cell carcinoma of kidney, reserve cell carcinoma, carcinomasarcomatodes, schneiderian carcinoma, scirrhous carcinoma, carcinomascroti, signet-ring cell carcinoma, carcinoma simplex, small-cellcarcinoma, solanoid carcinoma, spheroidal cell carcinoma, spindle cellcarcinoma, carcinoma spongiosum, squamous carcinoma, squamous cellcarcinoma, string carcinoma, carcinoma telangiectaticum, carcinomatelangiectodes, transitional cell carcinoma, carcinoma tuberosum,tuberous carcinoma, verrucous carcinoma, or carcinoma viflosum.

Additional cancers that can be treated include, e.g., Leukemia,Hodgkin's Disease, Non-Hodgkin's Lymphoma, multiple myeloma,neuroblastoma, breast cancer, ovarian cancer, lung cancer,rhabdomyosarcoma, primary thrombocytosis, primary macroglobulinemia,small-cell lung tumors, primary brain tumors, stomach cancer, coloncancer, malignant pancreatic insulanoma, malignant carcinoid, urinarybladder cancer, premalignant skin lesions, testicular cancer, lymphomas,thyroid cancer, papillary thyroid cancer, neuroblastoma, neuroendocrinecancer, esophageal cancer, genitourinary tract cancer, malignanthypercalcemia, cervical cancer, endometrial cancer, adrenal corticalcancer, prostate cancer, Müllerian cancer, ovarian cancer, peritonealcancer, fallopian tube cancer, or uterine papillary serous carcinoma.

2. Combination Therapy

The disclosure further includes a polynucleotide, e.g., mRNA, encoding atethered IL-12 polypeptide or uses thereof as a combination therapy,i.e., with any other anti-cancer agent in combination.

In certain embodiments, the disclosure is directed to a polynucleotide,e.g., mRNA, encoding a tethered IL-12 polypeptide in combination withone or more anti-cancer agents or uses of the polynucleotide incombination with one or more anti-cancer agents to the subject. In oneembodiment, the combination therapy can be a combination of thepolynucleotide, e.g., mRNA, encoding IL-12 and one or more standardtherapy. In another embodiment, the methods of the disclosure includetwo additional anti-cancer agents, three additional agents, fouradditional agents, etc. The additional anti-cancer agents can be aprotein, e.g., an antibody, or a polynucleotide, e.g., mRNA. In someembodiments, the one or more anti-cancer agents are an mRNA. In certainembodiments, the one or more anti-cancer agents are a polynucleotideencoding a tumor antigen. In certain embodiments, the one or moreanti-cancer agents are an mRNA encoding a tumor antigen. In otherembodiments, the one or more anti-cancer agents are not a tumor antigenor an mRNA encoding a tumor antigen. In other embodiments, the one ormore anti-cancer agents are a protein, e.g., an antibody.

In some embodiments, the one or more anti-cancer agents are an approvedagent by the United States Food and Drug Administration. In otherembodiments, the one or more anti-cancer agents are a pre-approved agentby the United States Food and Drug Administration.

One skilled in the art would also appreciate that alternativeembodiments of the present disclosure include a combination therapy of apolynucleotide, e.g., mRNA, encoding a tethered IL-12 polypeptide andany other agents, e.g., an anti-PD-1 antibody, an anti-PD-L1 antibody,and/or an anti-CTLA-4 antibody. For example, the present disclosureencompasses combination therapy of (i) a polynucleotide (e.g., mRNA)encoding a tethered IL-12 polypeptide and a protein comprising ananti-PD-1 antibody or an anti-PD-L1 antibody; or (iii) a polynucleotide(e.g., mRNA) encoding a tethered IL-12 polypeptide and a second proteincomprising an anti-CTLA-4 antibody.

In other embodiments, the additional agents can be formulated togetherwith the polynucleotide encoding a tethered IL-12 polypeptide, e.g.,mRNA, or separately. Moreover, even when formulated separately, theadditional agents can be administered concurrently with thepolynucleotide, e.g., mRNA, encoding a tethered IL-12 polypeptide orsequentially. In one embodiment, the polynucleotide, e.g., mRNA,encoding a tethered IL-12 polypeptide is administered prior to thesecond agent. In another embodiment, the polynucleotide, e.g., mRNA,encoding a tethered IL-12 polypeptide is administered after the secondagent.

In certain embodiments, the additional agents, e.g., any antibodydisclosed herein, are also administered intratumorally. In otherembodiments, the second agents, e.g., any antibody disclosed herein, areadministered via different routes, e.g., intravenously, subcutaneously,intraperitoneally, etc.

In some aspects, the subject for the present methods or compositions hasbeen treated with one or more standard of care therapies. In otheraspects, the subject for the present methods or compositions has notbeen responsive to one or more standard of care therapies or anti-cancertherapies. In one aspect, the subject has been previously treated withan IL-12 protein or an IL-12 DNA gene therapy. In another aspect, thesubject is treated with an anti-PD-1 antagonist prior to thepolynucleotide of the present disclosure. In another aspect, the subjecthas been treated with a monoclonal antibody that binds to PD-1 prior tothe polynucleotide of the present disclosure. In another aspect, thesubject has been treated with an anti-PD-1 monoclonal antibody therapyprior to the polynucleotide of the present methods or compositions.

In recent years, the introduction of immune checkpoint inhibitors fortherapeutic purposes has revolutionized cancer treatment. Of interestare therapies featuring combinations of checkpoint inhibitors with othercostimulatory or inhibitory molecules.

T cell regulation, i.e., activation or inhibition, is mediated viaco-stimulatory or co-inhibitory signals. This interaction is exerted vialigand/receptor interaction. T cells harbor a myriad of both activatingreceptors, such as OX40, and inhibitory receptors (i.e., immunecheckpoints) such as programmed death receptor 1 (PD-1) or cytotoxic Tlymphocyte-associated protein 4 (CTLA-4) (Mellman et al. 2011 Nature;480:480-489). Activation of these immune checkpoints results in T celldeactivation and commandeering these pathways by tumor cells contributesto their successful immune escape.

Immune checkpoint inhibitors such as pembrolizumab or nivolumab, whichtarget the interaction between programmed death receptor 1/programmeddeath ligand 1 (PD-1/PD-L1) and PD-L2, have been recently approved forthe treatment of various malignancies and are currently beinginvestigated in clinical trials for cancers including melanoma, head andneck squamous cell carcinoma (HNSCC). Data available from these trialsindicate substantial activity accompanied by a favorable safety andtoxicity profile in these patient populations.

For example, checkpoint inhibitors have been tested in clinical trialsfor the treatment of melanoma. In particular, phase III clinical trialshave revealed that therapies such as ipilimumab and pembrolizumab, whichtarget the CTLA-4 and PD-1 immune checkpoints, respectively, have raisedthe three-year survival of patients with melanoma to ˜70%, and overallsurvival (>5 years) to ˜30%.

Likewise, checkpoint inhibitors have been tested in clinical trials forthe treatment of head and neck cancer. In preclinical studies, it hadbeen shown that that 45-80% of HNSCC tumors express programmed deathligand 1 (PD-L1) (Zandberg et al. (2014) Oral Oncol. 50:627-632).Currently there are dozens of clinical trials evaluating the efficacyand safety of immune checkpoint inhibitors as monotherapy or incombination regimens in HNSCC. For example, clinical trials with PD 1,PD-L1, and CTLA-4 inhibitors are being tested in HNSCC. Data that thePD-1 antibody pembrolizumab might be effective in metastatic/recurrent(R/M) HNSCC patients were generated in the phase 1b Keynote-012 phaseI/II trial (Cheng. ASCO 2015, oral presentation). More recently the dataof the randomized CheckMate-141 phase III clinical trial were presented(Gillison. AACR 2016, oral presentation). This study investigated theefficacy of the monoclonal PD-1 antibody nivolumab given every 2 weeksin platinum-refractory R/M HNSCC patients. The study was stopped earlydue to superiority of the nivolumab arm of the study.

In one aspect, the subject has been previously treated with a PD-1antagonist prior to the polynucleotide of the present disclosure. Inanother aspect, the subject has been treated with a monoclonal antibodythat binds to PD-1 prior to the polynucleotide of the presentdisclosure. In another aspect, the subject has been treated with ananti-PD-1 monoclonal antibody therapy prior to the polynucleotide of thepresent methods or compositions. In other aspects, the anti-PD-1monoclonal antibody therapy comprises nivolumab, pembrolizumab,pidilizumab, or any combination thereof. In another aspect, the subjecthas been treated with a monoclonal antibody that binds to PD-L1 prior tothe polynucleotide of the present disclosure. In another aspect, thesubject has been treated with an anti-PD-L1 monoclonal antibody therapyprior to the polynucleotide of the present methods or compositions. Inother aspects, the anti-PD-L1 monoclonal antibody therapy comprisesdurvalumab, avelumab, MEDI473, BMS-936559, aezolizumab, or anycombination thereof.

In some aspects, the subject has been treated with a CTLA-4 antagonistprior to treatment with the compositions of present disclosure. Inanother aspect, the subject has been previously treated with amonoclonal antibody that binds to CTLA-4 prior to the compositions ofthe present disclosure. In another aspect, the subject has been treatedwith an anti-CTLA-4 monoclonal antibody prior to the polynucleotide ofthe present disclosure. In other aspects, the anti-CTLA-4 antibodytherapy comprises ipilimumab or tremelimumab.

In some aspects, the disclosure is directed to a method of treatingcancer and/or a method of immunotherapy in a subject in need thereofcomprising administering to the subject (e.g., intratumorally,intraperitoneally, or intravenously) the polynucleotide (e.g., RNA,e.g., mRNA) encoding a tethered IL-12 polypeptide in combination with aPD-L1 antagonist, e.g., an antibody or antigen-binding portion thereofthat specifically binds to PD-L1, e.g., an anti-PD-L1 monoclonalantibody, e.g., an anti-PD-L1 monoclonal antibody comprises Durvalumab,Avelumab, MEDI473, BMS-936559, Atezolizumab, or any combination thereof.

In certain embodiments, the anti-PD-L1 antibody useful for thedisclosure is MSB0010718C (also called Avelumab; See US 2014/0341917) orBMS-936559 (formerly 12A4 or MDX-1105) (see, e.g., U.S. Pat. No.7,943,743; WO 2013/173223). In other embodiments, the anti-PD-L1antibody is MPDL3280A (also known as RG7446) (see, e.g., Herbst et al.(2013) J Clin Oncol 31 (suppl): 3000. Abstract; U.S. Pat. No.8,217,149), MEDI4736 (also called Durvalumab; Khleif (2013) In:Proceedings from the European Cancer Congress 2013; Sep. 27-Oct. 1,2013; Amsterdam, The Netherlands.

In some aspects, the disclosure is directed to a method of treatingcancer and/or a method of immunotherapy in a subject in need thereofcomprising administering to the subject (e.g., intratumorally,intraperitoneally, or intravenously) the polynucleotide (e.g., RNA,e.g., mRNA) encoding a tethered IL-12 polypeptide, in combination with aPD-1 antagonist, e.g., an antibody or antigen-binding portion thereofthat specifically binds to PD-1, e.g., an anti-PD-1 monoclonal antibody.

In one embodiment, the anti-PD-1 antibody (or an antigen-binding portionthereof) useful for the disclosure is pembrolizumab. Pembrolizumab (alsoknown as KEYTRUIDA®, lambrolizumab, and MK-3475) is a humanizedmonoclonal IgG4 antibody directed against human cell surface receptorPD-1 (programmed death-1 or programmed cell death-1). Pembrolizumab isdescribed, for example, in U.S. Pat. No. 8,900,587; see alsohttp://www.cancer.gov/drugdictionary?cdrid=695789 (last accessed: Dec.14, 2014). Pembrolizumab has been approved by the FDA for the treatmentof relapsed or refractory melanoma and advanced NSCLC.

In another embodiment, the anti-PD-1 antibody useful for the disclosureis nivolumab. Nivolumab (also known as “OPDIVO®”; formerly designated5C4, BMS-936558, MDX-1106, or ONO-4538) is a fully human IgG4 (S228P)PD-1 immune checkpoint inhibitor antibody that selectively preventsinteraction with PD-1 ligands (PD-L1 and PD-L2), thereby blocking thedown-regulation of antitumor T-cell functions (U.S. Pat. No. 8,008,449;Wang et al., 2014 Cancer Immunol Res. 2(9):846-56). Nivolumab has shownactivity in a variety of advanced solid tumors including renal cellcarcinoma (renal adenocarcinoma, or hypernephroma), melanoma, andnon-small cell lung cancer (NSCLC) (Topalian et al., 2012a; Topalian etal., 2014; Drake et al., 2013; WO 2013/173223.

In other embodiments, the anti-PD-1 antibody is MEDI0680 (formerlyAMP-514), which is a monoclonal antibody against the PD-1 receptor.MEDI0680 is described, for example, in U.S. Pat. No. 8,609,089B2 or inhttp://www.cancer.gov/drugdictionary?cdrid=756047 (last accessed Dec.14, 2014).

In certain embodiments, the anti-PD-1 antibody is BGB-A317, which is ahumanized monoclonal antibody. BGB-A317 is described in U.S. Publ. No.2015/0079109.

In certain embodiments, a PD-1 antagonist is AMP-224, which is a B7-DCFc fusion protein. AMP-224 is discussed in U.S. Publ. No. 2013/0017199or inhttp://www.cancer.gov/publications/dictionaries/cancer-drug?cdrid=700595(last accessed Jul. 8, 2015).

In other embodiments, the disclosure includes a method of treatingcancer and/or a method of immunotherapy in a subject in need thereofcomprising administering to the subject (e.g., intratumorally,intraperitoneally, or intravenously) the polynucleotide (e.g., RNA,e.g., mRNA) encoding a tethered IL-12 polypeptide together with anantibody or an antigen binding portion thereof that specifically bindsto PD-1, e.g., an anti-PD-1 monoclonal antibody, e.g., an anti-PD-1monoclonal antibody comprises Nivolumab, Pembrolizumab, Pidilizumab, orany combination thereof.

In other aspects, the disclosure is directed to a method of treatingcancer and/or a method of immunotherapy in a subject in need thereofcomprising administering to the subject (e.g., intratumorally,intraperitoneally, or intravenously) the polynucleotide (e.g., RNA,e.g., mRNA) encoding a tethered IL-12 polypeptide in combination with aCTLA-4 antagonist, e.g., an antibody or antigen-binding portion thereofthat specifically binds to CTLA-4, e.g., an anti-CTLA-4 monoclonalantibody, e.g., an anti-CTLA-4 monoclonal antibody comprises Ipilimumabor Tremelimumab, or any combination thereof.

An exemplary clinical anti-CTLA-4 antibody is the human mAb 10D1 (nowknown as ipilimumab and marketed as YERVOY®) as disclosed in U.S. Pat.No. 6,984,720. Another anti-CTLA-4 antibody useful for the presentmethods is tremelimumab (also known as CP-675,206). Tremelimumab ishuman IgG2 monoclonal anti-CTLA-4 antibody. Tremelimumab is described inWO/2012/122444, U.S. Publ. No. 2012/263677, or WO Publ. No. 2007/113648A2.

In some embodiments, the compositions disclosed herein comprise (i) apolynucleotide, e.g., mRNA, encoding a tethered IL-12 polypeptide and(ii) a polynucleotide, e.g., mRNA, encoding an antibody or an antigenbinding portion thereof which specifically binds to CTLA-4 in a singleformulation.

3. Interleukin-12 (IL-12)

IL-12 (also shown as IL12) is a pleiotropic cytokine, the actions ofwhich create an interconnection between innate and adaptive immunity.IL-12 functions primarily as a 70 kDa heterodimeric protein consistingof two disulfide-linked p35 and p40 subunits. The precursor form of theIL-12 p40 subunit (NM 002187; P29460; also referred to as IL-12B,natural killer cell stimulatory factor 2, cytotoxic lymphocytematuration factor 2) is 328 amino acids in length, while its mature formis 306 amino acids long. The precursor form of the IL-12 p35 subunit(NM_000882; P29459; also referred to as IL-12A, natural killer cellstimulatory factor 1, cytotoxic lymphocyte maturation factor 1) is 219amino acids in length and the mature form is 197 amino acids long. Id.The genes for the IL-12 p35 and p40 subunits reside on differentchromosomes and are regulated independently of each other. Gately, M Ket al., Annu Rev Immunol. 16: 495-521 (1998). Many different immunecells (e.g., dendritic cells, macrophages, monocytes, neutrophils, and Bcells) produce IL-12 upon antigenic stimuli. The active IL-12heterodimer is formed following protein synthesis. Id.

IL-12 is composed of a bundle of four alpha helices. It is aheterodimeric cytokine encoded by two separate genes, IL-12A (p35) andIL-12B (p40). The active heterodimer (referred to as ‘p′70’), and ahomodimer of p40 are formed following protein synthesis.

In some embodiments, the tethered IL-12 polypeptide of the presentdisclosure comprises IL-12A. In some embodiments, the tethered IL-12polypeptide of the present disclosure comprises IL-12B. In someembodiments, the tethered IL-12 polypeptide of the present disclosurecomprises both IL-12A and IL-12B.

In some embodiments, IL-12B is located N-terminal to IL-12A in thetethered IL-12 polypeptide of the present disclosure. In someembodiments, IL-12A is located N-terminal to IL-12B in the tetheredIL-12 polypeptide of the present disclosure. The phrase “locatedN-terminal to” indicates location in a polypeptide with respect to othersequences in the polypeptide in relation to the N-terminus of thepolypeptide. For example, IL-12B that is “N-terminal to” IL-12A meansthat IL-12B is located closer to the N-terminus of the tethered IL-12polypeptide than IL-12A.

In some embodiments, the tethered IL-12 polypeptide of the presentdisclosure comprises a single polypeptide chain comprising IL-12B andIL-12A, which are fused directly to one another or are linked to oneanother by a linker (referred to herein as an “subunit linker”).Non-limiting examples of linkers are disclosed elsewhere herein.

In some embodiments, the tethered IL-12 polypeptide of the disclosurecomprises IL-12A and/or IL-12B that is a variant, that is a functionalfragment, or that contains a substitution, an insertion and/or anaddition, a deletion, and/or a covalent modification with respect to awild-type IL-12A or IL-12B sequence. In some embodiments, sequence tags(such as epitope tags, e.g., a V5 tag) or amino acids, can be added tothe sequences encoded by the polynucleotides of the disclosure (e.g., atthe N-terminal or C-terminal ends), e.g., for localization. In someembodiments, amino acid residues located at the carboxy, amino terminal,or internal regions of a polypeptide of the disclosure can optionally bedeleted providing for fragments.

In some embodiments, the tethered IL-12 polypeptide encoded by apolynucleotide of the disclosure (e.g., a RNA, e.g., an mRNA) comprisesa substitutional variant of an IL-12A and/or IL-12B sequence, which cancomprise one, two, three or more than three substitutions. In someembodiments, the substitutional variant can comprise one or moreconservative amino acids substitutions. In other embodiments, thevariant is an insertional variant. In other embodiments, the variant isa deletional variant.

As recognized by those skilled in the art, IL-12 protein fragments,functional protein domains, variants, and homologous proteins(orthologs) are also considered to be within the scope of the IL-12polypeptides of the disclosure. Nonlimiting examples of polypeptidesencoded by the polynucleotides of the disclosure are set forth in SEQ IDNOs: 1, 3, 45, 48 and 49. For example, SEQ ID NOs: 1, 3, and 45 providethe amino acid sequence of human wild type IL-12.

4. Membrane Domains

The tethered IL-12 polypeptides of the disclosure comprise a membranedomain that tethers (i.e., anchors) the IL-12 polypeptide to a cellmembrane (e.g., a transmembrane domain). In some embodiments, thetethered IL-12 polypeptides comprise a transmembrane domain. In someembodiments, the tethered IL-12 polypeptides comprise a transmembranedomain, and optionally an intracellular domain. In some embodiments, thetethered IL-12 polypeptides comprise a transmembrane domain and anintracellular domain.

In some embodiments, the membrane domain is from an integral membraneprotein.

Integral membrane proteins can include, for example, integral polytopicproteins that contain a single-pass or multi-pass transmembrane domainthat tethers the protein to a cell surface, including domains withhydrophobic α-helical or β-barrel (i.e., β-sheet) structures. Theamino-terminus (i.e., N-terminus) of Type I integral membrane proteinsis located in the extracellular space, while the carboxy-terminus (i.e.,C-terminus) of Type II integral membrane proteins is located in theintracellular space.

In some embodiments, a tethered IL-12 polypeptide of the disclosurecomprises a transmembrane domain from an integral polytopic protein. Insome embodiments, a tethered IL-12 polypeptide of the disclosurecomprises a transmembrane domain from a Type I integral membraneprotein. In some embodiments, a tethered IL-12 polypeptide comprises atransmembrane domain from a Type II integral membrane protein.

In some embodiments, the transmembrane domain comprises an intracellulardomain (i.e., a domain that is localized to the intracellular space of acell, e.g., a domain that is localized to the cytoplasm of a cell). Insome embodiments, an intracellular domain has been removed from thetransmembrane domain. In some embodiments, the transmembrane domaincomprises a membrane domain without an intracellular domain.

Integral membrane proteins can also include, for example, integralmonotopic proteins that contain a membrane domain that does not span theentire cell membrane but that tethers the protein to a cell surface.

In some embodiments, a tethered IL-12 polypeptide of the disclosurecomprises a membrane domain from an integral monotopic protein.

In some embodiments, the membrane domain is from a Cluster ofDifferentiation (CD) protein, CD8, CD80, CD4, a receptor,Platelet-Derived Growth Factor Receptor (PDGF-R), Interleukin-6 Receptor(IL-6R), transferrin receptor, Tumor Necrosis Factor (TNF) receptor,erythropoietin (EPO) receptor, a T Cell Receptor (TCR), TCR β-chain, aFc receptor, FcγRII, FcεRI, an interferon receptor, type I interferonreceptor, a growth factor, Stem Cell Factor (SCF), TNF-α, B7-1,Asialoglycoprotein, c-erbB-2, ICAM-1, an immunoglobulin, an IgG, an IgM,a viral glycoprotein, rabies virus glycoprotein, respiratory syncytialvirus glycoprotein G (RSVG), vesicular stomatis virus glycoprotein(VSVG), a viral hemagglutinin (HA), influenza HA, vaccinia virus HA, orany combination thereof.

In some embodiments, the membrane domain is selected from the groupconsisting of: a CD8 transmembrane domain, a PDGF-R transmembranedomain, a CD80 transmembrane domain, and any combination thereof.

Exemplary transmembrane domains are set forth in SEQ ID NOs: 101-103.

In one embodiment, a membrane domain comprises a transmembrane domain ofT-cell surface glycoprotein CD8 alpha chain (also known as CD8A orT-lymphocyte differentiation antigen T8/Leu-2), e.g., a transmembrane ofUniProtKB—P01732. In some embodiments, the polynucleotide, e.g., mRNA,encoding a tethered IL-12, comprises a nucleotide sequence encoding aCD8 transmembrane polypeptide. In some embodiments, the polynucleotide,e.g., mRNA, encoding a tethered IL-12, comprises a nucleotide sequenceencoding a CD8 transmembrane polypeptide as set forth in SEQ ID NO: 101.

In another embodiment, a membrane domain comprises a transmembranedomain of platelet-derived growth factor receptor beta (EC:2.7.10.1)(also known as PDGF-R-beta, PDGFR-beta, beta platelet-derived growthfactor receptor, beta-type platelet-derived growth factor receptor,CD140 antigen-like family member B, platelet-derived growth factorreceptor 1, PDGFR-1, or CD140b), e.g., a transmembrane domain ofUniProtKB—P09619. In some embodiments, the polynucleotide, e.g., mRNA,encoding a tethered IL-12, comprises a nucleotide sequence encoding aPDGFR-beta transmembrane polypeptide. In some embodiments, thepolynucleotide, e.g., mRNA, encoding a tethered IL-12, comprises anucleotide sequence encoding a PDGFR-beta transmembrane polypeptide asset forth in SEQ ID NO: 102.

In other embodiments, a membrane domain comprises a transmembrane domainof T-lymphocyte activation antigen CD80 (also known as activation B7-1antigen, BB1, CTLA-4 counter-receptor B7.1, or B7), e.g., atransmembrane domain of UniProtKB—P33681. In some embodiments, thepolynucleotide, e.g., mRNA, encoding a tethered IL-12, comprises anucleotide sequence encoding a CD80 transmembrane polypeptide. In someembodiments, the polynucleotide, e.g., mRNA, encoding a tethered IL-12,comprises a nucleotide sequence encoding a CD80 transmembranepolypeptide as set forth in SEQ ID NO: 103.

In some embodiments, the membrane domain in the tethered IL-12polypeptide comprises an amino acid sequence at least about 50%, atleast about 60%, at least about 70%, at least about 80%, at least about90%, at least about 95%, at least about 99%, or about 100% identical toSEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, or any combinationthereof. In other embodiments, the membrane domain in the tethered IL-12polypeptide comprises an amino acid sequence set forth in SEQ ID NO:101, SEQ ID NO: 102, or SEQ ID NO: 103 without one amino acid, two aminoacids, three amino acids, four amino acids, five amino acids, six aminoacids, seven amino acids, eight amino acids, or nine amino acids at theN terminus of SEQ ID NO: 101, SEQ ID NO: 102, or SEQ ID NO: 103 and/orwithout one amino acid, two amino acids, three amino acids, four aminoacids, five amino acids, six amino acids, seven amino acids, eight aminoacids, or nine amino acids at the C terminus of SEQ ID NO: 101, SEQ IDNO: 102, or SEQ ID NO: 103. In certain embodiments, the membrane domainin the tethered IL-12 polypeptide comprises SEQ ID NO: 101, SEQ ID NO:102, SEQ ID NO: 103, or any combination thereof with one amino acidsubstitution, two amino acid substitutions, three amino acidsubstitutions, four amino acid substitutions, five amino acidsubstitutions, six amino acid substitutions, seven amino acidsubstitutions, eight amino acid substitutions, or nine amino acidsubstitutions, wherein the membrane domain is capable of tethering theIL-12 polypeptide on a cell membrane.

In some embodiments, the membrane domain comprises a transmembranedomain and an intracellular domain. In some embodiments, anintracellular domain is any oligopeptide or polypeptide known to act asa transmission signal in a cell. In some embodiments, the membranedomain comprises an intracellular domain to stabilize the tethered IL-12polypeptide.

Intracellular domains useful in the methods and compositions of thepresent disclosure include at least those derived from any of thepolypeptides in which transmembrane domains are derived, as describedsupra. For example, suitable intracellular domains include, but are notlimited to, an intracellular domain derived from CD80, PDGFR, or anycombination thereof.

In some embodiments, a membrane domain comprises an intracellular domainof platelet-derived growth factor receptor beta (EC:2.7.10.1) (alsoknown as PDGF-R-beta, PDGFR-beta, beta platelet-derived growth factorreceptor, beta-type platelet-derived growth factor receptor, CD140antigen-like family member B, platelet-derived growth factor receptor 1,PDGFR-1, or CD140b), e.g., an intracellular domain of UniProtKB—P09619.In some embodiments, the polynucleotide, e.g., mRNA, encoding a tetheredIL-12, comprises a nucleotide sequence encoding a PDGFR-betaintracellular polypeptide. In some embodiments, the polynucleotide,e.g., mRNA, encoding a tethered IL-12, comprises a nucleotide sequenceencoding a PDGFR-beta intracellular polypeptide as set forth in SEQ IDNO: 226.

In some embodiments, a membrane domain comprises a truncatedintracellular domain of PDGFR-beta. In some embodiments, a truncatedintracellular domain of PDGFR-beta stabilizes the tethered IL-12polypeptide compared to the wild-type PDGFR-beta intracellular domain.In some embodiments, the polynucleotide, e.g., mRNA, encoding a tetheredIL-12, comprises a nucleotide sequence encoding a truncated PDGFR-betaintracellular polypeptide. In some embodiments, the polynucleotide,e.g., mRNA, encoding a tethered IL-12, comprises a nucleotide sequenceencoding a truncated PDGFR-beta intracellular polypeptide as set forthin SEQ ID NO: 227. In some embodiments, the polynucleotide, e.g., mRNA,encoding a tethered IL-12, comprises a nucleotide sequence encoding atruncated PDGFR-beta intracellular polypeptide as set forth in SEQ IDNO: 228.

In other embodiments, a membrane domain comprises an intracellulardomain of T-lymphocyte activation antigen CD80 (also known as activationB7-1 antigen, BB1, CTLA-4 counter-receptor B7.1, or B7), e.g., anintracellular domain of UniProtKB—P33681. In some embodiments, thepolynucleotide, e.g., mRNA, encoding a tethered IL-12, comprises anucleotide sequence encoding a CD80 intracellular polypeptide. In someembodiments, the polynucleotide, e.g., mRNA, encoding a tethered IL-12,comprises a nucleotide sequence encoding a CD80 intracellularpolypeptide as set forth in SEQ ID NO: 225.

In some embodiments, the tethered IL-12 polypeptides described hereincomprise a membrane domain comprising a transmembrane domain and anintracellular domain derived from the same polypeptide (i.e.,homologous). In some embodiments, the tethered IL-12 polypeptidesdescribed herein comprise a membrane domain comprising a CD80transmembrane domain and CD80 intracellular domain. In some embodiments,the tethered IL-12 polypeptide described herein comprise a membranedomain comprising a PDGFR-beta transmembrane domain and PDGFR-betaintracellular domain. In some embodiments, the tethered IL-12polypeptides described herein comprise a membrane domain comprising atransmembrane domain and an intracellular domain derived from differentpolypeptides (i.e., heterologous) (e.g., a CD80 transmembrane domain anda PDGFR-beta intracellular domain; a CD8 transmembrane domain and a CD80intracellular domain; a CD8 transmembrane domain and a PDGFR-betatransmembrane domain; or a PDGFR-beta transmembrane domain and a CD80intracellular domain).

In some embodiments, the membrane domain (e.g., transmembrane domain,and optional intracellular domain) in the tethered IL-12 polypeptide islocated C-terminal to any IL-12 amino acid sequence (i.e., any aminoacid sequence of IL-12A, IL-12B, or both IL-12A and IL-12B when both arepresent in the tethered IL-12 polypeptide). The phrase “locatedC-terminal to” indicates location in a polypeptide with respect to othersequences in the polypeptide in relation to the C-terminus of thepolypeptide. A membrane domain (e.g., transmembrane domain, and optionalintracellular domain) that is “C-terminal to” any IL-12 amino acidsequences means that the membrane domain is located closer to theC-terminus of the tethered IL-12 polypeptide than any IL-12 amino acidsequences.

In some embodiments, the membrane domain (e.g., transmembrane domain,and optional intracellular domain) in the tethered IL-12 polypeptide isfrom a Type I integral membrane protein and is located C-terminal to anyIL-12 amino acid sequence.

In some embodiments, the membrane domain (e.g., transmembrane domain,and optional intracellular domain) in the tethered IL-12 polypeptide islocated N-terminal to the IL-12 polypeptide. A membrane domain that is“N-terminal to” any IL-12 amino acid sequences means that the membranedomain is located closer to the N-terminus of the tethered IL-12polypeptide than any IL-12 amino acid sequences.

In some embodiments, the membrane domain (e.g., transmembrane domain,and optional intracellular domain) in the tethered IL-12 polypeptide isfrom a Type II integral membrane protein and is located N-terminal toany IL-12 amino acid sequence.

In some embodiments, the membrane domain (e.g., transmembrane domain,and optional intracellular domain) in the tethered IL-12 polypeptide islinked to the IL-12 polypeptide by a linker, which is referred to hereinas a “membrane domain linker” or a “transmembrane domain linker” whenthe membrane domain is a transmembrane domain, and optionally anintracellular domain. Non-limiting examples of linkers are disclosedelsewhere herein. In some embodiments, the membrane domain in thetethered IL-12 polypeptide is fused directly to the IL-12 polypeptide.

5. Polynucleotides and Open Reading Frames (ORFs)

In certain aspects, the disclosure provides polynucleotides (e.g., aRNA, e.g., an mRNA) that comprise a nucleotide sequence encoding atethered IL-12 polypeptide.

In one aspect, the disclosure provides polynucleotides (e.g., a RNA,e.g., an mRNA) comprising a nucleic acid sequence encoding an IL-12polypeptide and a nucleic acid sequence encoding a membrane domain(e.g., transmembrane domain, and optional intracellular domain), whereinthe nucleic acid sequence encoding the membrane domain is fused directlyto the nucleic acid sequence encoding the IL-12 polypeptide or is linkedto the nucleic acid sequence encoding the IL-12 polypeptide by a nucleicacid sequence encoding a linker (membrane domain linker).

In some aspects, the disclosure provides polynucleotides (e.g., a RNA,e.g., an mRNA) comprising a nucleic acid sequence encoding an IL-12polypeptide, and a nucleic acid sequence encoding a transmembranedomain, wherein the nucleic acid sequence encoding the transmembranedomain is fused directly to the nucleic acid sequence encoding the IL-12polypeptide or is linked to the nucleic acid sequence encoding the IL-12polypeptide by a nucleic acid sequence encoding a linker (membranedomain linker).

In some aspects, the disclosure provides polynucleotides (e.g., a RNA,e.g., an mRNA) comprising a nucleic acid sequence encoding an IL-12polypeptide, a nucleic acid sequence encoding a transmembrane domain,and a nucleic acid encoding an intracellular domain, wherein the nucleicacid sequence encoding the transmembrane domain is fused directly to thenucleic acid sequence encoding the IL-12 polypeptide or is linked to thenucleic acid sequence encoding the IL-12 polypeptide by a nucleic acidsequence encoding a linker (membrane domain linker), and wherein thenucleic acid encoding the intracellular domain is fused directly to thenucleic acid sequence encoding the transmembrane domain, or is linked tothe nucleic acid sequence encoding the transmembrane domain by a nucleicacid sequence encoding a linker.

The skilled artisan will appreciate that it is possible to directly fusedomains within encoded chimeric proteins. As such it is possible todirectly fuse domains by omitting linker sequences (e.g., omittingflexible linker-encoding sequences). For example, there are describedherein constructs in which a membrane domain (e.g., a transmembranedomain, and optional intracellular domain) is directly fused to aninterleukin domain or chain. As is shown herein, however, suchconstructions can lead to diminished interleukin activity as compared,for example, to corresponding constructs having flexible linkersequences included.

In some embodiments, the IL-12 polypeptide comprises an IL-12 p40subunit (IL-12B), an IL-12A p35 subunit (IL-12A), or both.

In some embodiments, the nucleic acid sequence encoding the membranedomain (e.g., transmembrane domain, and optional intracellular domain)is located at the 3′ terminus of the nucleic acid sequence encoding theIL-12 polypeptide or at the 3′ terminus of the nucleic acid sequenceencoding the membrane domain linker.

In some embodiments, the nucleic acid sequence encoding the membranedomain (e.g., transmembrane domain, and optional intracellular domain)is located at the 5′ terminus of the nucleic acid sequence encoding theIL-12 polypeptide or at the 5′ terminus of the nucleic acid sequenceencoding the membrane domain linker.

In some embodiments, the polynucleotide comprises an open reading frame(ORF) comprising a nucleic acid sequence encoding IL-12B, a nucleic acidsequence encoding IL-12A, and a nucleic acid sequence encoding amembrane domain (e.g., transmembrane domain, and optional intracellulardomain), wherein the ORF optionally comprises a nucleic acid sequenceencoding a linker (subunit linker) that links the IL-12B and the IL-12A.

In some embodiments, the nucleic acid sequence encoding the IL-12B islocated at the 5′ terminus of the nucleic acid sequence encoding theIL-12A or at the 5′ terminus of the nucleic acid sequence encoding thesubunit linker.

In some embodiments, the nucleic acid sequence encoding the IL-12B islocated at the 3′ terminus of the nucleic acid sequence encoding theIL-12A or at the 3′ terminus of the nucleic acid sequence encoding thesubunit linker.

In some embodiments, the nucleic acid sequence encoding the membranedomain (e.g., transmembrane domain, and optional intracellular domain)is located at the 3′ terminus of the nucleic acid sequence encoding theIL-12A or at the 3′ terminus of the membrane domain linker.

In some embodiments, the nucleic acid sequence encoding the membranedomain (e.g., transmembrane domain, and optional intracellular domain)is located at the 5′ terminus of the nucleic acid sequence encoding theIL-12B or at the 5′ terminus of the membrane domain linker.

In some embodiments, the IL-12 polypeptide comprises an IL-12Bpolypeptide (i.e., IL-12B) selected from:

(a) the full-length IL-12B polypeptide (e.g., having the same oressentially the same length as wild-type IL-12B);

(b) a functional fragment of the wild-type IL-12B polypeptide (e.g., atruncated (e.g., deletion of carboxy, amino terminal, or internalregions) sequence shorter than an IL-12B wild-type; but still retainingIL-12B enzymatic activity);

(c) a variant thereof (e.g., full length or truncated IL-12B proteins inwhich one or more amino acids have been replaced, e.g., variants thatretain all or most of the IL-12B activity of the polypeptide withrespect to the wild type IL-12B polypeptide (such as, e.g., V33I, V298F,or any other natural or artificial variants known in the art); and

(d) a fusion protein comprising (i) a full length IL-12B wild-type, afunctional fragment or a variant thereof, and (ii) a heterologousprotein.

In some embodiments, the IL-12 polypeptide comprises an IL-12Apolypeptide (i.e., IL-12A) selected from:

(a) the full-length IL-12A polypeptide (e.g., having the same oressentially the same length as wild-type IL-12A);

(b) a functional fragment of the wild-type IL-12A polypeptide (e.g., atruncated (e.g., deletion of carboxy, amino terminal, or internalregions) sequence shorter than an IL-12A wild-type; but still retainingIL-12A enzymatic activity);

(c) a variant thereof (e.g., full length or truncated IL-12A proteins inwhich one or more amino acids have been replaced, e.g., variants thatretain all or most of the IL-12A activity of the polypeptide withrespect to the wtIL-12A polypeptide (such as natural or artificialvariants known in the art); and

(d) a fusion protein comprising (i) a full length IL-12A wild-type, afunctional fragment or a variant thereof, and (ii) a heterologousprotein.

In some embodiments, the membrane domain is selected from:

(a) the full-length membrane domain (e.g., having the same oressentially the same length as the wild-type membrane domain);

(b) a functional fragment of the wild-type membrane domain (e.g., atruncated (e.g., deletion of carboxy, amino terminal, or internalregions) sequence shorter than a membrane domain wild-type; but stillretaining the ability of a membrane domain to tether a polypeptide to acell membrane);

(c) a variant thereof (e.g., full length or truncated membrane domainproteins in which one or more amino acids have been replaced, e.g.,variants that retain the ability of a membrane domain to tether apolypeptide to a cell membrane); and

(d) a fusion protein comprising (i) a full length membrane domainwild-type, a functional fragment or a variant thereof, and (ii) aheterologous protein.

In certain embodiments, the polynucleotide (e.g., a RNA, e.g., an mRNA)of the disclosure comprises a nucleic acid sequence encoding a mammalianIL-12 polypeptide and/or membrane domain (e.g., a polypeptide comprisingmammalian IL-12A, mammalian IL-12B, or both mammalian IL-12A andmammalian IL-12B), such as a human IL-12 polypeptide and/or membranedomain (e.g., a polypeptide comprising human IL-12A, human IL-12B, orboth human IL-12A and human IL-12B), including a functional fragment ora variant thereof.

In some embodiments, the polynucleotide (e.g., a RNA, e.g., an mRNA) ofthe disclosure increases IL-12B and/or IL-12A protein expression levelsand/or detectable IL-12 enzymatic activity levels in cells whenintroduced in those cells, e.g., by at least 10%, at least 15%, at least20%, at least 25%, at least 30%, at least 35%, at least 40%, at least45%, at least 50%, at least 55%, at least 60%, at least 65%, at least70%, at least 75%, at least 80%, at least 85%, at least 90%, at least95%, or at least 100%, compared to IL-12B and/or IL-12A proteinexpression levels and/or detectable IL-12 enzymatic activity levels inthe cells prior to the administration of the polynucleotide of thedisclosure. IL-12B and/or IL-12A protein expression levels and/or IL-12enzymatic activity can be measured according to methods known in theart. In some embodiments, the polynucleotide is introduced to the cellsin vitro. In some embodiments, the polynucleotide is introduced to thecells in vivo.

In some embodiments, the polynucleotides (e.g., a RNA, e.g., an mRNA) ofthe disclosure comprise a nucleotide sequence (e.g., an ORF comprising anucleotide sequence) that encodes a wild-type human IL-12B and/orIL-12A.

In some embodiments, the polynucleotide (e.g., a RNA, e.g., an mRNA) ofthe disclosure comprises a codon optimized nucleic acid sequence,wherein the open reading frame (ORF) of the codon optimized nucleicsequence is derived from a wild-type IL-12A and/or IL-12B sequence.

In some embodiments, the polynucleotides (e.g., a RNA, e.g., an mRNA) ofthe disclosure comprise a nucleotide sequence encoding IL-12B and/orIL-12A having the full length sequence of human IL-12B and/or IL-12A(i.e., including the initiator methionine and the signal peptides). Inmature human IL-12B and/or IL-12A, the initiator methionine and/orsignal peptides can be removed to yield a “mature IL-12B” and/or “matureIL-12A” comprising amino acid residues of SEQ ID NO: 1 and SEQ ID NO: 3,respectively. SEQ ID NO: 1 corresponds to amino acids 23 to 328 of SEQID NO: 48, and SEQ ID NO: 3 corresponds to amino acids 336 to 532 of SEQID NO: 48. The teachings of the present disclosure directed to the fullsequence of human IL-12B and/or IL-12A are also applicable to the matureform of human IL-12B and/or IL-12A lacking the initiator methionineand/or the signal peptide. Thus, in some embodiments, thepolynucleotides (e.g., a RNA, e.g., an mRNA) of the disclosure comprisea nucleotide sequence encoding IL-12B and/or IL-12A having the maturesequence of human IL-12B and/or IL-12A. In some embodiments, thepolynucleotide (e.g., a RNA, e.g., an mRNA) of the disclosure comprisinga nucleotide sequence encoding IL-12B and/or IL-12A is sequenceoptimized.

In some embodiments, the polynucleotides (e.g., a RNA, e.g., an mRNA) ofthe disclosure comprise a nucleotide sequence (e.g., an ORF comprising anucleotide sequence) encoding a tethered IL-12 polypeptide comprising amutant IL-12B and/or IL-12A polypeptide. In some embodiments, thepolynucleotides of the disclosure comprise an ORF comprising anucleotide sequence encoding an IL-12B and/or IL-12A polypeptide thatcomprises at least one point mutation in the IL-12B and/or IL-12Asequence and retains IL-12B and/or IL-12A enzymatic activity. In someembodiments, the mutant IL-12B and/or IL-12A polypeptide has an IL-12Band/or IL-12A activity which is at least 10%, at least 15%, at least20%, at least 25%, at least 30%, at least 35%, at least 40%, at least45%, at least 50%, at least 55%, at least 60%, at least 65%, at least70%, at least 75%, at least 80%, at least 85%, at least 90%, at least95%, or at least 100% of the IL-12B and/or IL-12A activity of thecorresponding wild-type IL-12B and/or IL-12A (i.e., the same IL-12Band/or IL-12A but without the mutation(s)). In some embodiments, thepolynucleotide (e.g., a RNA, e.g., an mRNA) of the disclosure comprisingan ORF encoding a mutant IL-12B and/or IL-12A polypeptide is sequenceoptimized.

In some embodiments, the polynucleotide (e.g., a RNA, e.g., an mRNA) ofthe disclosure comprises a nucleotide sequence (e.g., an ORF comprisinga nucleotide sequence) that encodes an IL-12B and/or IL-12A polypeptidewith mutations that do not alter IL-12B and/or IL-12A enzymaticactivity. Such mutant IL-12B and/or IL-12A polypeptides can be referredto as function-neutral. In some embodiments, the polynucleotidecomprises an ORF comprising a nucleotide sequence that encodes a mutantIL-12B and/or IL-12A polypeptide comprising one or more function-neutralpoint mutations.

In some embodiments, the mutant IL-12B and/or IL-12A polypeptide hashigher IL-12B and/or IL-12A enzymatic activity than the correspondingwild-type IL-12B and/or IL-12A. In some embodiments, the mutant IL-12Band/or IL-12A polypeptide has an IL-12B and/or IL-12A activity that isat least 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 55%, atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, or at least 100% higher than theactivity of the corresponding wild-type IL-12B and/or IL-12A (i.e., thesame IL-12B and/or IL-12A but without the mutation(s)).

In some embodiments, the polynucleotides (e.g., a RNA, e.g., an mRNA) ofthe disclosure comprise a nucleotide sequence (e.g., an ORF comprising anucleotide sequence) encoding a functional IL-12B and/or IL-12Afragment, e.g., where one or more fragments correspond to a polypeptidesubsequence of a wild type IL-12B and/or IL-12A polypeptide and retainIL-12B and/or IL-12A enzymatic activity. In some embodiments, the IL-12Band/or IL-12A fragment has an IL-12B and/or IL-12A activity which is atleast 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 55%, atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, or at least 100% of the IL-12activity of the corresponding full length IL-12B and/or IL-12A. In someembodiments, the polynucleotides (e.g., a RNA, e.g., an mRNA) of thedisclosure comprising an ORF comprising a nucleotide sequence encoding afunctional IL-12B and/or IL-12A fragment is sequence optimized.

In some embodiments, the polynucleotide (e.g., a RNA, e.g., an mRNA) ofthe disclosure comprises a nucleotide sequence (e.g., an ORF comprisinga nucleotide sequence) encoding an IL-12B and/or IL-12A fragment thathas higher IL-12B and/or IL-12A enzymatic activity than thecorresponding full length IL-12B and/or IL-12A. Thus, in someembodiments the IL-12B and/or IL-12A fragment has an IL-12B and/orIL-12A activity which is at least 10%, at least 15%, at least 20%, atleast 25%, at least 30%, at least 35%, at least 40%, at least 45%, atleast 50%, at least 55%, at least 60%, at least 65%, at least 70%, atleast 75%, at least 80%, at least 85%, at least 90%, at least 95%, or atleast 100% higher than the IL-12B and/or IL-12A activity of thecorresponding full length IL-12B and/or IL-12A.

In some embodiments, the polynucleotide (e.g., a RNA, e.g., an mRNA) ofthe disclosure comprises a nucleotide sequence (e.g., an ORF comprisinga nucleotide sequence) encoding an IL-12B and/or IL-12A fragment that isat least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%,15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24% or 25% shorter thanwild-type IL-12B and/or IL-12A.

In other embodiments, the polynucleotide (e.g., a RNA, e.g., an mRNA) ofthe disclosure comprises a nucleotide sequence (e.g., an ORF comprisinga nucleotide sequence) encoding IL-12B, which has an amino acid sequenceat least about 80%, at least about 90%, at least about 95%, at least96%, at least 97%, at least 98%, at least 99%, or 100% identical toamino acids 23 to 328 of SEQ ID NO: 48, and wherein the amino acidsequence has IL-12B activity.

In other embodiments, the polynucleotide (e.g., a RNA, e.g., an mRNA) ofthe disclosure comprises a nucleotide sequence (e.g., an ORF comprisinga nucleotide sequence) encoding IL-12A, which has an amino acid sequenceat least about 80%, at least about 90%, at least about 95%, at least96%, at least 97%, at least 98%, at least 99%, or 100% identical toamino acids 336 to 532 of SEQ ID NO:48, and wherein the amino acidsequence has IL-12A activity.

In other embodiments, the polynucleotide (e.g., a RNA, e.g., an mRNA) ofthe disclosure comprises a nucleotide sequence (e.g., an ORF comprisinga nucleotide sequence) encoding IL-12B, which has:

(i) at least 70%, at least 75%, at least 76%, at least 77%, at least78%, at least 79%, at least 80%, at least 81%, at least 82%, at least83%, at least 84%, at least 85%, at least 86%, at least 87%, at least88%, at least 89%, at least 90%, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least98%, at least 99%, or 100% sequence identity to nucleotides 67-984 ofhIL12AB_001 to hIL12AB_003 (SEQ ID NOs: 5 to 7), hIL12AB_005 tohIL12AB_040 (SEQ ID NO: 9 to 44), or hIL12AB_041 (SEQ ID NO: 220); or

(ii) at least 70%, at least 75%, at least 76%, at least 77%, at least78%, at least 79%, at least 80%, at least 81%, at least 82%, at least83%, at least 84%, at least 85%, at least 86%, at least 87%, at least88%, at least 89%, at least 90%, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least98%, at least 99%, or 100% sequence identity to nucleotides 70-987 ofhIL12AB_004 (SEQ ID NO: 8).

In other embodiments, the polynucleotide (e.g., a RNA, e.g., an mRNA) ofthe disclosure comprises a nucleotide sequence (e.g., an ORF comprisinga nucleotide sequence) encoding IL-12A, which has:

(i) at least 70%, at least 75%, at least 76%, at least 77%, at least78%, at least 79%, at least 80%, at least 81%, at least 82%, at least83%, at least 84%, at least 85%, at least 86%, at least 87%, at least88%, at least 89%, at least 90%, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least98%, at least 99%, or 100% sequence identity to nucleotides 1006-1596 ofhIL12AB_001 to hIL12AB_003 (SEQ ID NOs: 5 to 7), hIL12AB_005 tohIL12AB_040 (SEQ ID NO: 9 to 44), or hIL12AB_041 (SEQ ID NO: 220); or

(ii) at least 70%, at least 75%, at least 76%, at least 77%, at least78%, at least 79%, at least 80%, at least 81%, at least 82%, at least83%, at least 84%, at least 85%, at least 86%, at least 87%, at least88%, at least 89%, at least 90%, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least98%, at least 99%, or 100% sequence identity to nucleotides 1009-1599 ofhIL12AB_004 (SEQ ID NO: 4).

In some embodiments, the polynucleotide (e.g., a RNA, e.g., an mRNA) ofthe disclosure comprises a nucleotide sequence (e.g., an ORF) encodingan IL-12B-IL-12A fusion polypeptide (e.g., the wild-type sequence,functional fragment, or variant thereof), wherein the nucleotidesequence has at least 60%, at least 65%, at least 70%, at least 71%, atleast 72%, at least 73%, at least 74%, at least 75%, at least 76%, atleast 77%, at least 78%, at least 79%, at least 80%, at least 81%, atleast 82%, at least 83%, at least 84%, at least 85%, at least 86%, atleast 87%, at least 88%, at least 89%, at least 90%, at least 91%, atleast 92%, at least 93%, at least 94%, at least 95%, at least 96%, atleast 97%, at least 98%, at least 99%, or 100% sequence identity to asequence selected from the group consisting of SEQ ID NOs: 5 to 44.

In some embodiments, the polynucleotide (e.g., a RNA, e.g., an mRNA) ofthe disclosure comprises a nucleotide sequence (e.g., an ORF comprisinga nucleotide sequence) encoding an IL-12B-IL-12A fusion polypeptide(e.g., the wild-type sequence, functional fragment, or variant thereof),wherein the nucleotide sequence has 70% to 100%, 75% to 100%, 80% to100%, 85% to 100%, 70% to 95%, 80% to 95%, 70% to 85%, 75% to 90%, 80%to 95%, 70% to 75%, 75% to 80%, 80% to 85%, 85% to 90%, 90% to 95%, or95% to 100%, sequence identity to a sequence selected from the groupconsisting of SEQ ID NOs: 5 to 44 or 220.

In some embodiments, the polynucleotide (e.g., a RNA, e.g., an mRNA)encoding the IL-12 polypeptide comprises a nucleotide sequence at least60%, at least 65%, at least 70%, at least 71%, at least 72%, at least73%, at least 74%, at least 75%, at least 76%, at least 77%, at least78%, at least 79%, at least 80%, at least 81%, at least 82%, at least83%, at least 84%, at least 85%, at least 86%, at least 87%, at least88%, at least 89%, at least 90%, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least98%, at least 99%, or 100% sequence identity to SEQ ID NO: 6(hIL12AB_002).

In some embodiments, the polynucleotide (e.g., a RNA, e.g., an mRNA)encoding the IL-12 polypeptide comprises a nucleotide sequence at least60%, at least 65%, at least 70%, at least 71%, at least 72%, at least73%, at least 74%, at least 75%, at least 76%, at least 77%, at least78%, at least 79%, at least 80%, at least 81%, at least 82%, at least83%, at least 84%, at least 85%, at least 86%, at least 87%, at least88%, at least 89%, at least 90%, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least98%, at least 99%, or 100% sequence identity to SEQ ID NO: 220(hIL12AB_041).

In some embodiments, the polynucleotide (e.g., a RNA, e.g., an mRNA) ofthe disclosure comprises from about 900 to about 100,000 nucleotides(e.g., from 900 to 1,000, from 900 to 1,100, from 900 to 1,200, from 900to 1,300, from 900 to 1,400, from 900 to 1,500, from 1,000 to 1,100,from 1,000 to 1,100, from 1,000 to 1,200, from 1,000 to 1,300, from1,000 to 1,400, from 1,000 to 1,500, from 1,083 to 1,200, from 1,083 to1,400, from 1,083 to 1,600, from 1,083 to 1,800, from 1,083 to 2,000,from 1,083 to 3,000, from 1,083 to 5,000, from 1,083 to 7,000, from1,083 to 10,000, from 1,083 to 25,000, from 1,083 to 50,000, from 1,083to 70,000, or from 1,083 to 100,000).

In some embodiments, the polynucleotide of the disclosure (e.g., a RNA,e.g., an mRNA) comprises a nucleotide sequence encoding an IL-12B-IL-12Afusion polypeptide (e.g., an ORF comprising a nucleotide sequenceencoding IL-12B and a nucleotide encoding IL-12A; e.g., the wild-typesequence, functional fragment, or variant thereof encoding the IL-12Band/or IL-12A), wherein the length of the nucleotide sequence is atleast 500 nucleotides in length (e.g., at least or greater than about500, 600, 700, 80, 900, 1,000, 1,050, 1,083, 1,100, 1,200, 1,300, 1,400,1,500, 1,600, 1,700, 1,800, 1,900, 2,000, 2,100, 2,200, 2,300, 2,400,2,500, 2,600, 2,700, 2,800, 2,900, 3,000, 3,100, 3,200, 3,300, 3,400,3,500, 3,600, 3,700, 3,800, 3,900, 4,000, 4,100, 4,200, 4,300, 4,400,4,500, 4,600, 4,700, 4,800, 4,900, 5,000, 5,100, 5,200, 5,300, 5,400,5,500, 5,600, 5,700, 5,800, 5,900, 6,000, 7,000, 8,000, 9,000, 10,000,20,000, 30,000, 40,000, 50,000, 60,000, 70,000, 80,000, 90,000 or up toand including 100,000 nucleotides).

In one embodiment, the nucleic acid sequence encoding the IL-12B and thenucleic acid sequence encoding the IL-12A are fused directly to oneanother. In another embodiment, the nucleic acid sequence encoding theIL-12B and the nucleic acid sequence (encoding the IL-12A are linked bya nucleic acid sequence encoding a subunit linker.

In one embodiment, the nucleic acid sequence encoding the membranedomain is fused directly to the nucleic acid sequence encoding the IL-12polypeptide comprising IL-12A, IL-12B, or IL-12A and IL-12B. In anotherembodiment, the nucleic acid sequence encoding the membrane domain islinked to the nucleic acid sequence encoding the IL-12 polypeptidecomprising IL-12A, IL-12B, or IL-12A and IL-12B by a nucleic acidsequence encoding a membrane domain linker.

In some embodiments, the polynucleotide of the disclosure (e.g., a RNA,e.g., an mRNA) further comprises at least one nucleic acid sequence thatis noncoding, e.g., a miRNA binding site.

In some embodiments, the polynucleotide of the disclosure (e.g., a RNA,e.g., an mRNA) is single stranded or double stranded.

In some embodiments, the polynucleotide of the disclosure is DNA or RNA.In some embodiments, the polynucleotide of the disclosure is RNA. Insome embodiments, the polynucleotide of the disclosure is, or functionsas, a messenger RNA (mRNA). In some embodiments, the mRNA comprises anucleotide sequence that encodes IL-12B and/or IL-12A and a nucleic acidsequence encoding a membrane domain, and is capable of being translatedto produce the encoded tethered IL-12 polypeptide in vitro, in vivo, insitu or ex vivo.

In some embodiments, the polynucleotide of the disclosure (e.g., a RNA,e.g., an mRNA) comprises a sequence-optimized nucleotide sequenceencoding IL-12B and/or IL-12A (e.g., the wild-type sequence, functionalfragment, or variant thereof encoding the IL-12B and/or the IL-12A)and/or a sequence-optimized nucleotide sequence encoding the membranedomain, wherein the polynucleotide comprises at least one chemicallymodified nucleobase, e.g., 5-methoxyuracil. In some embodiments, thepolynucleotide further comprises a miRNA binding site, e.g., a miRNAbinding site that binds to miR-122. In some embodiments, thepolynucleotide disclosed herein is formulated with a delivery agent,e.g., a compound having Formula (I).

The polynucleotides (e.g., a RNA, e.g., an mRNA) of the disclosure canalso comprise nucleotide sequences that encode additional features thatfacilitate trafficking of the encoded polypeptides to therapeuticallyrelevant sites. One such feature that aids in protein trafficking is thesignal sequence, or targeting sequence. The peptides encoded by thesesignal sequences are known by a variety of names, including targetingpeptides, transit peptides, and signal peptides. In some embodiments,the polynucleotide (e.g., a RNA, e.g., an mRNA) comprises a nucleotidesequence (e.g., an ORF comprising a nucleotide sequence) that encodes asignal peptide operably linked a nucleotide sequence that encodes anIL-12B and/or IL-12A polypeptide described herein.

In some embodiments, the “signal sequence” or “signal peptide” is apolynucleotide or polypeptide, respectively, which is from about 9 to200 nucleotides (3-70 amino acids) in length that, optionally, isincorporated at the 5′ (or N-terminus) of the coding region or thepolypeptide, respectively. Some signal peptides are cleaved from theprotein, for example by a signal peptidase after the proteins aretransported to the desired site.

In some embodiments, the polynucleotide (e.g., a RNA, e.g., an mRNA) ofthe disclosure comprises a nucleotide sequence encoding an IL-12B and/orIL-12A polypeptide, wherein the nucleotide sequence further comprises a5′ nucleic acid sequence encoding a native signal peptide. In anotherembodiment, the polynucleotide of the disclosure comprises a nucleotidesequence encoding an IL-12B and/or IL-12A polypeptide, wherein thenucleotide sequence lacks the nucleic acid sequence encoding a nativesignal peptide.

In some embodiments, the polynucleotide (e.g., a RNA, e.g., an mRNA) ofthe disclosure comprises a nucleotide sequence encoding an IL-12B and/orIL-12A polypeptide, wherein the nucleotide sequence further comprises a5′ nucleic acid sequence encoding a heterologous signal peptide.

In some embodiments, the polynucleotide (e.g., a RNA, e.g., an mRNA)further comprises a nucleic acid sequence encoding a signal peptide thatis located at the 5′ terminus of the nucleotide sequence encoding theIL-12B.

In some embodiments, the nucleotide sequence encoding the IL-12Bcomprises a nucleic acid sequence encoding a signal peptide.

In some embodiments, the signal peptide comprises a sequence at leastabout 80%, at least about 90%, at least about 95%, at least 96%, atleast 97%, at least 98%, at least 99%, or 100% identical to amino acids1 to 22 of SEQ ID NO: 48.

In some aspects, the disclosure provides mRNA comprising an open readingframe (ORF), wherein the ORF comprises a nucleotide sequence encodingfrom 5′ to 3′:

-   -   5′-[IL-12B]-[L1]-[IL-12A]-[L2]-[MD]-3′, wherein    -   IL-12B is a human IL-12 p40 subunit polypeptide,    -   L1 is a first peptide linker,    -   IL-12A is a human IL-12 p35 subunit polypeptide,    -   L2 is a second peptide linker,    -   MD is a membrane domain comprising a transmembrane domain, and        optionally an intracellular domain.

In some embodiments, the polynucleotide (e.g., mRNA) encoding a tetheredIL-12 polypeptide comprises an open reading frame (ORF) encoding IL-12B,IL-12A, and a CD8 transmembrane domain, wherein the ORF optionallyencodes a linker that links the IL-12B and the IL-12A, and wherein theORF optionally encodes a linker that links the CD8 transmembrane domainand the IL-12B.

In some embodiments, the polynucleotide (e.g., mRNA) encoding a tetheredIL-12 polypeptide comprises an open reading frame (ORF) encoding IL-12B,IL-12A, and a CD8 transmembrane domain, wherein the ORF optionallyencodes a linker that links the IL-12B and the IL-12A, and wherein theORF optionally encodes a linker that links the CD8 transmembrane domainand the IL-12A.

In some embodiments, the polynucleotide (e.g., mRNA) encoding a tetheredIL-12 polypeptide comprises an open reading frame (ORF) encoding IL-12B,IL-12A, and a CD80 transmembrane domain, wherein the ORF optionallyencodes a linker that links the IL-12B and the IL-12A, and wherein theORF optionally encodes a linker that links the CD80 transmembrane domainand the IL-12B.

In some embodiments, the polynucleotide (e.g., mRNA) encoding a tetheredIL-12 polypeptide comprises an open reading frame (ORF) encoding IL-12B,IL-12A, and a CD80 transmembrane domain, wherein the ORF optionallyencodes a linker that links the IL-12B and the IL-12A, and wherein theORF optionally encodes a linker that links the CD80 transmembrane domainand the IL-12A.

In some embodiments, the polynucleotide (e.g., mRNA) encoding a tetheredIL-12 polypeptide comprises an open reading frame (ORF) encoding IL-12B,IL-12A, and a PDGFR-beta transmembrane domain, wherein the ORFoptionally encodes a linker that links the IL-12B and the IL-12A, andwherein the ORF optionally encodes a linker that links the PDGFR-betatransmembrane domain and the IL-12B.

In some embodiments, the polynucleotide (e.g., mRNA) encoding a tetheredIL-12 polypeptide comprises an open reading frame (ORF) encoding IL-12B,IL-12A, and a PDGFR-beta transmembrane domain, wherein the ORFoptionally encodes a linker that links the IL-12B and the IL-12A, andwherein the ORF optionally encodes a linker that links the PDGFR-betatransmembrane domain and the IL-12A.

In some embodiments, the polynucleotide (e.g., mRNA) encoding a tetheredIL-12 polypeptide comprises an open reading frame (ORF) encoding IL-12B,IL-12A, a CD80 transmembrane domain and intracellular domain, whereinthe ORF optionally encodes a linker that links the IL-12B and theIL-12A, and wherein the ORF optionally encodes a linker that links theCD80 transmembrane domain and the IL-12B.

In some embodiments, the polynucleotide (e.g., mRNA) encoding a tetheredIL-12 polypeptide comprises an open reading frame (ORF) encoding IL-12B,IL-12A, a CD80 transmembrane domain and intracellular domain, whereinthe ORF optionally encodes a linker that links the IL-12B and theIL-12A, and wherein the ORF optionally encodes a linker that links theCD80 transmembrane domain and the IL-12A.

In some embodiments, the polynucleotide (e.g., mRNA) encoding a tetheredIL-12 polypeptide comprises an open reading frame (ORF) encoding IL-12B,IL-12A, a PDGFR-beta transmembrane domain and intracellular domain,wherein the ORF optionally encodes a linker that links the IL-12B andthe IL-12A, and wherein the ORF optionally encodes a linker that linksthe CD80 transmembrane domain and the IL-12B.

In some embodiments, the polynucleotide (e.g., mRNA) encoding a tetheredIL-12 polypeptide comprises an open reading frame (ORF) encoding IL-12B,IL-12A, a PDGFR-beta transmembrane domain and intracellular domain,wherein the ORF optionally encodes a linker that links the IL-12B andthe IL-12A, and wherein the ORF optionally encodes a linker that linksthe CD80 transmembrane domain and the IL-12A.

In some embodiments, the polynucleotide (e.g., mRNA) encoding a tetheredIL-12 polypeptide comprises an open reading frame (ORF) encoding IL-12B,IL-12A, a PDGFR-beta transmembrane domain and truncated intracellulardomain, wherein the ORF optionally encodes a linker that links theIL-12B and the IL-12A, and wherein the ORF optionally encodes a linkerthat links the CD80 transmembrane domain and the IL-12B.

In some embodiments, the polynucleotide (e.g., mRNA) encoding a tetheredIL-12 polypeptide comprises an open reading frame (ORF) encoding IL-12B,IL-12A, a PDGFR-beta transmembrane domain and truncated intracellulardomain, wherein the ORF optionally encodes a linker that links theIL-12B and the IL-12A, and wherein the ORF optionally encodes a linkerthat links the CD80 transmembrane domain and the IL-12A.

In some embodiments, the polynucleotide (e.g., mRNA) encoding a tetheredIL-12 polypeptide comprises an open reading frame (ORF) encoding anIL-12 polypeptide comprising the amino acid sequence set forth in SEQ IDNO: 48, a linker comprising the amino acid sequence set forth in SEQ IDNO: 229, and a CD8 transmembrane domain comprising the amino acidsequence set forth in SEQ ID NO: 101.

In some embodiments, the polynucleotide (e.g., mRNA) encoding a tetheredIL-12 polypeptide comprises an open reading frame (ORF) encoding anIL-12 polypeptide comprising the amino acid sequence set forth in SEQ IDNO: 48, a linker comprising the amino acid sequence set forth in SEQ IDNO: 229, and a CD80 transmembrane domain comprising the amino acidsequence set forth in SEQ ID NO: 103.

In some embodiments, the polynucleotide (e.g., mRNA) encoding a tetheredIL-12 polypeptide comprises an open reading frame (ORF) encoding anIL-12 polypeptide comprising the amino acid sequence set forth in SEQ IDNO: 48, a linker comprising the amino acid sequence set forth in SEQ IDNO: 229, and a PDGFR-beta transmembrane domain comprising the amino acidsequence set forth in SEQ ID NO: 102.

In some embodiments, the polynucleotide (e.g., mRNA) encoding a tetheredIL-12 polypeptide comprises an open reading frame (ORF) encoding anIL-12 polypeptide comprising the amino acid sequence set forth in SEQ IDNO: 48, a linker comprising the amino acid sequence set forth in SEQ IDNO: 229, a CD80 transmembrane domain comprising the amino acid sequenceset forth in SEQ ID NO: 103, and a CD80 intracellular domain comprisingthe amino acid sequence set forth in SEQ ID NO: 225.

In some embodiments, the polynucleotide (e.g., mRNA) encoding a tetheredIL-12 polypeptide comprises an open reading frame (ORF) encoding anIL-12 polypeptide comprising the amino acid sequence set forth in SEQ IDNO: 48, a linker comprising the amino acid sequence set forth in SEQ IDNO: 229, a CD80 transmembrane domain comprising the amino acid sequenceset forth in SEQ ID NO: 103, and a truncated PDGFR-beta intracellulardomain comprising the amino acid sequence set forth in SEQ ID NO: 227.

In some embodiments, the polynucleotide (e.g., mRNA) encoding a tetheredIL-12 polypeptide comprises an open reading frame (ORF) encoding anIL-12 polypeptide comprising the amino acid sequence set forth in SEQ IDNO: 48, a linker comprising the amino acid sequence set forth in SEQ IDNO: 229, a CD80 transmembrane domain comprising the amino acid sequenceset forth in SEQ ID NO: 103, and a truncated PDGFR-beta intracellulardomain comprising the amino acid sequence set forth in SEQ ID NO: 228.

In some embodiments, the polynucleotide encoding a tethered IL-12polypeptide is an mRNA comprising an open reading frame (ORF),comprising SEQ ID NO: 221, SEQ ID NO: 376, and SEQ ID NO: 377. In someembodiments, the polynucleotide encoding a tethered IL-12 polypeptide isan mRNA comprising a 5′UTR comprising SEQ ID NO: 287; an open readingframe (ORF), comprising SEQ ID NO: 221, SEQ ID NO: 376, and SEQ ID NO:377; and a 3′UTR comprising SEQ ID NO: 283.

In some embodiments, the polynucleotide encoding a tethered IL-12polypeptide is an mRNA comprising an open reading frame (ORF) comprisingSEQ ID NO: 221, SEQ ID NO: 376, and SEQ ID NO: 378. In some embodiments,the polynucleotide encoding a tethered IL-12 polypeptide is an mRNAcomprising a 5′UTR comprising SEQ ID NO: 287; an open reading frame(ORF) comprising SEQ ID NO: 221, SEQ ID NO: 376, and SEQ ID NO: 378; anda 3′UTR comprising SEQ ID NO: 283.

In some embodiments, the polynucleotide encoding a tethered IL-12polypeptide is an mRNA comprising an open reading frame (ORF) comprisingSEQ ID NO: 221, SEQ ID NO: 376, and SEQ ID NO: 250. In some embodiments,the polynucleotide encoding a tethered IL-12 polypeptide is an mRNAcomprising a 5′UTR comprising SEQ ID NO: 287; an open reading frame(ORF) comprising SEQ ID NO: 221, SEQ ID NO: 376, and SEQ ID NO: 250; anda 3′UTR comprising SEQ ID NO: 283.

In some embodiments, the polynucleotide encoding a tethered IL-12polypeptide is an mRNA comprising an open reading frame (ORF) comprisingSEQ ID NO: 221, SEQ ID NO: 376, SEQ ID NO: 378 and SEQ ID NO: 379. Insome embodiments, the polynucleotide encoding a tethered IL-12polypeptide is an mRNA comprising a 5′UTR comprising SEQ ID NO: 287; anopen reading frame (ORF) comprising SEQ ID NO: 221, SEQ ID NO: 376, SEQID NO: 378 and SEQ ID NO: 379; and a 3′UTR comprising SEQ ID NO: 283.

In some embodiments, the polynucleotide encoding a tethered IL-12polypeptide is an mRNA comprising an open reading frame (ORF) comprisingSEQ ID NO: 221, SEQ ID NO: 376, SEQ ID NO: 250 and SEQ ID NO: 251. Insome embodiments, the polynucleotide encoding a tethered IL-12polypeptide is an mRNA comprising a 5′UTR comprising SEQ ID NO: 287; anopen reading frame (ORF) comprising SEQ ID NO: 221, SEQ ID NO: 376, SEQID NO: 250 and SEQ ID NO: 325; and a 3′UTR comprising SEQ ID NO: 283.

In some embodiments, the polynucleotide encoding a tethered IL-12polypeptide is an mRNA comprising an open reading frame (ORF) comprisingSEQ ID NO: 221, SEQ ID NO: 376, SEQ ID NO: 250 and SEQ ID NO: 280. Insome embodiments, the polynucleotide encoding a tethered IL-12polypeptide is an mRNA comprising a 5′UTR comprising SEQ ID NO: 287; anopen reading frame (ORF) comprising SEQ ID NO: 221, SEQ ID NO: 376, SEQID NO: 250 and SEQ ID NO: 280; and a 3′UTR comprising SEQ ID NO: 283.

6. Chimeric Proteins

In some embodiments, the polynucleotide of the disclosure (e.g., a RNA,e.g., an mRNA) can comprise more than one nucleic acid sequence encodingone or more polypeptides. In some embodiments, polynucleotides of thedisclosure comprise an a nucleic acid sequence encoding an IL-12polypeptide comprising IL-12A and/or IL-12B and a nucleic acid sequenceencoding a membrane domain, including functional fragments or a variantsthereof (i.e., any or all of the IL-12A, IL-12B, or the membrane domaincan be a functional fragment or variant). In some embodiments, thepolynucleotide of the disclosure can comprise a nucleic acid sequenceencoding IL-12B, a nucleic acid sequence encoding IL-12A, and a nucleicacid sequence encoding a membrane domain, including functional fragmentsor a variant thereof (i.e., any or all of the IL-12A, IL-12B, or themembrane domain can be a functional fragment or variant). In someembodiments, the polynucleotide of the disclosure can comprise one ormore additional nucleic acid sequences expressing one or more additionalpolypeptides of interest (e.g., one or more additional nucleic acidsequences encoding one or more polypeptides heterologous to IL-12). Inone embodiment, the additional polypeptide of interest can be fused tothe IL-12B polypeptide directly or by a linker. In another embodiment,the additional polypeptide of interest can be fused to the IL-12Apolypeptide directly or by a linker. In other embodiments, theadditional polypeptide of interest can be fused to both the IL-12Bpolypeptide and the IL-12A polypeptide directly or by a linker. In otherembodiments, the first additional polypeptide of interest is fused tothe IL-12A polypeptide directly or by a linker, and the secondadditional polypeptide of interest is fused to the IL-12B polypeptidedirectly or by a linker. In some embodiments, two or more additionalpolypeptides of interest can be genetically fused, i.e., two or moreadditional polypeptides of interest can be encoded by the same ORF. Insome embodiments, the polynucleotide can comprise a nucleic acidsequence encoding a linker (e.g., a G4S peptide linker or another linkerknown in the art) between two or more additional polypeptides ofinterest.

In some embodiments, the polynucleotide of the disclosure (e.g., a RNA,e.g., an mRNA) can comprise a nucleic acid sequence (e.g., one or morenucleic acid sequences) encoding an IL-12B polypeptide, IL-12Apolypeptide, both IL-12B and IL-12A polypeptides, a nucleic acidsequence encoding a membrane domain, and one or more additional nucleicacid sequences encoding one or more additional polypeptides of interest.

7. Linker

In one aspect, the membrane domain can be fused directly to the IL-12polypeptide of the present disclosure comprising IL-12A, IL-12B, and/orIL-12A and IL-12B or can be linked to the IL-12 polypeptide by a linker(referred to herein as the “membrane domain linker” or a “transmembranedomain linker” when the membrane domain is a transmembrane domain). Inanother aspect, IL-12B and IL-12A in an IL-12 polypeptide can be fuseddirectly to one another or can be linked to one another by a linker(referred to herein as the “subunit linker”). In other embodiments, theIL-12B and/or IL-12A can be fused directly to a heterologous polypeptideor can be linked to the heterologous polypeptide by a linker (referredto herein as the “heterologous polypeptide linker.”). In otherembodiments, the membrane domain can be fused directly to a heterologouspolypeptide or can be linked to the heterologous polypeptide byheterologous polypeptide linker. Suitable linkers can be a polypeptide(or peptide) moiety or a non-polypeptide moiety. In some embodiments,the linker is a peptide linker, including from one amino acid to about200 amino acids. In some embodiments, the linker comprises at least 1,at least 2, at least 3, at least 4, at least 5, at least 6, at least 7,at least 8, at least 9, at least 10, at least 11, at least 12, at least13, at least 14, at least 15, at least 16, at least 17, at least 18, atleast 19, at least 20, at least 21, at least 22, at least 23, at least24, at least 25, at least 26, at least 27, at least 28, at least 29, atleast 30, at least 31, at least 32, at least 33, at least 34, at least35, at least 36, at least 37, at least 38, at least 39, or at least 40amino acids.

Without being bound in theory, it is believed that incorporation oflinker-encoding sequences, (in particular, flexible linker-encodingsequences) between sequences (e.g., ORF sequences) encoding functionaldomains (e.g., interleukin chains, transmembrane domains, etc.) providea certain degree of movement or interaction between domains, thusimproving functionality of the mRNA-encoded chimeric (e.g., “tethered”)interleukins of the disclosure. Flexible linkers are generally composedof small, non-polar (e.g., Gly) or polar (e.g., Ser) amino acids. Thesmall size of these amino acids provides flexibility, and allows formobility of the connected functional domains. Preferred flexible linkersencoded by sequences within the mRNAs of the disclosure have sequencesconsisting primarily of stretches of Gly and Ser residues (“GS” linker).An exemplary flexible linker has the sequence of(Gly-Gly-Gly-Gly-Ser)_(n). By adjusting the copy number “n”, the lengthof this GS linker can be optimized to achieve appropriate separation ofthe functional domains, or to maintain necessary inter-domaininteractions. In some embodiments, the membrane domain linker is ofsufficient length to prevent steric hindrance from the cell membrane.

In some embodiments, the linker can be a GS (Gly/Ser) linker, forexample, comprising (G_(n)S)_(m), wherein n is an integer from 1 to 100and m is an integer from 1 to 100. In some embodiments, the Gly/Serlinker comprises (G_(n)S)_(m) (SEQ ID NO: 193), wherein n is from 1 to20, e.g., 1, 2 3, 4, 5, 6, 7, 8, 9, 10, 15, or 20 and m is from 1 to 20,e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, or 20. In some embodiments, theGS linker can comprise (GGGGS)_(o) (SEQ ID NO: 194), wherein o is aninteger from 1 to 5. In some embodiments, the GS linker can compriseGGSGGGGSGG (SEQ ID NO: 195), GGSGGGGG (SEQ ID NO: 196), or GSGSGSGS (SEQID NO: 197). In some embodiments, the GS linker comprises GGGGGGS (SEQID NO: 214).

In some embodiments, the linker suitable for the disclosure can be aGly-rich linker, for example, comprising (Gly)_(p) (SEQ ID NO: 198),wherein p is an integer from 1 to 100, e.g., from 1 to 40. In someembodiments, a Gly-rich linker can comprise GGGGG (SEQ ID NO: 192),GGGGGG (SEQ ID NO: 217), GGGGGGG (SEQ ID NO: 218) or GGGGGGGG (SEQ IDNO: 219).

In some embodiments, the linker suitable for the disclosure can comprise(EAAAK)_(q) (SEQ ID NO: 199), wherein q is an integer from 1 to 100,e.g., from 1 to 20, e.g., from 1 to 5. In one embodiment, the linkersuitable for the disclosure can comprise (EAAAK)₃.

Further exemplary linkers include, but not limited to, GGGGSLVPRGSGGGGS(SEQ ID NO: 200), GSGSGS (SEQ ID NO: 201), GGGGSLVPRGSGGGG (SEQ ID NO:202), GGSGGHMGSGG (SEQ ID NO: 203), GGSGGSGGSGG (SEQ ID NO: 204), GGSGG(SEQ ID NO: 205), GSGSGSGS (SEQ ID NO: 206), GGGSEGGGSEGGGSEGGG (SEQ IDNO: 207), AAGAATAA (SEQ ID NO: 208), GGSSG (SEQ ID NO: 209), GSGGGTGGGSG(SEQ ID NO: 210), GSGSGSGSGGSG (SEQ ID NO: 211), GSGGSGSGGSGGSG (SEQ IDNO: 212), and GSGGSGGSGGSGGS (SEQ ID NO: 213).

The nucleotides encoding the linkers can be constructed to fuse thesequences of the present disclosure. Based on the RNA sequencesprovided, a person of ordinary skill in the art would understand thecorresponding DNA sequence (e.g., conversion of uracil to thymine).Likewise, based on the DNA sequences provided, a person of ordinaryskill in the art would understand the corresponding RNA sequence (e.g.,conversion of thymine to uracil).

8. Sequence Optimization of Nucleotide Sequence Encoding an IL-12Polypeptide

In some embodiments, the polynucleotide (e.g., a RNA, e.g., an mRNA) ofthe disclosure is sequence optimized. In some embodiments, thepolynucleotide (e.g., a RNA, e.g., an mRNA) of the disclosure comprisesa nucleotide sequence encoding an IL-12 polypeptide (i.e., an IL-12Band/or IL-12A polypeptide), a nucleotide sequence encoding a membranedomain, a nucleotide sequence encoding another polypeptide of interest,a 5′-UTR, a 3′-UTR, a miRNA, a nucleotide sequence encoding a linker(e.g., a membrane domain linker, a subunit linker, and/or a heterologouspolypeptide linker), or any combination thereof that is sequenceoptimized.

A sequence-optimized nucleotide sequence (e.g., an codon-optimized mRNAsequence encoding an IL-12B and/or IL-12A polypeptide) is a sequencecomprising at least one synonymous nucleobase substitution with respectto a reference sequence (e.g., a wild type nucleotide sequence encodingan IL-12B and/or IL-12A polypeptide).

A sequence-optimized nucleotide sequence can be partially or completelydifferent in sequence from the reference sequence. For example, areference sequence encoding polyserine uniformly encoded by TCT codonscan be sequence-optimized by having 100% of its nucleobases substituted(for each codon, T in position 1 replaced by A, C in position 2 replacedby G, and T in position 3 replaced by C) to yield a sequence encodingpolyserine which would be uniformly encoded by AGC codons. Thepercentage of sequence identity obtained from a global pairwisealignment between the reference polyserine nucleic acid sequence and thesequence-optimized polyserine nucleic acid sequence would be 0%.However, the protein products from both sequences would be 100%identical.

Some sequence optimization (also sometimes referred to codonoptimization) methods are known in the art (and discussed in more detailbelow) and can be useful to achieve one or more desired results. Theseresults can include, e.g., matching codon frequencies in certain tissuetargets and/or host organisms to ensure proper folding; biasing G/Ccontent to increase mRNA stability or reduce secondary structures;minimizing tandem repeat codons or base runs that can impair geneconstruction or expression; customizing transcriptional andtranslational control regions; inserting or removing protein traffickingsequences; removing/adding post translation modification sites in anencoded protein (e.g., glycosylation sites); adding, removing orshuffling protein domains; inserting or deleting restriction sites;modifying ribosome binding sites and mRNA degradation sites; adjustingtranslational rates to allow the various domains of the protein to foldproperly; and/or reducing or eliminating problem secondary structureswithin the polynucleotide. Sequence optimization tools, algorithms andservices are known in the art, non-limiting examples include servicesfrom GeneArt (Life Technologies), DNA2.0 (Menlo Park Calif.) and/orproprietary methods. Codon options for each amino acid are given inTable 1.

TABLE 1 Codon Options Single Letter Amino Acid Code Codon OptionsIsoleucine I ATT, ATC, ATA Leucine L CTT, CTC, CTA, CTG, TTA, TTG ValineV GTT, GTC, GTA, GTG Phenylalanine F TTT, TTC Methionine M ATG CysteineC TGT, TGC Alanine A GCT, GCC, GCA, GCG Glycine G GGT, GGC, GGA, GGGProline P CCT, CCC, CCA, CCG Threonine T ACT, ACC, ACA, ACG Serine sTCT, TCC, TCA, TCG, AGT, AGC Tyrosine Y TAT, TAC Tryptophan W TGGGlutamine Q CAA, CAG Asparagine N AAT, AAC Histidine H CAT, CACGlutamic acid E GAA, GAG Aspartic acid D GAT, GAC Lysine K AAA, AAGArginine R CGT, CGC, CGA, CGG, AGA, AGG Selenocysteine SecUGA in mRNA in presence of Selenocysteine insertion element (SECIS)Stop codons Stop TAA, TAG, TGA

In some embodiments, a polynucleotide (e.g., a RNA, e.g., an mRNA) ofthe disclosure comprises a sequence-optimized nucleotide sequenceencoding an IL-12 polypeptide, including a functional fragment or avariant thereof, or a sequence-optimized nucleotide sequence encoding amembrane domain, including a functional fragment or a variant thereof,wherein the IL-12 polypeptide and/or membrane domain encoded by thesequence-optimized nucleotide sequence has improved properties (e.g.,compared to an IL-12 polypeptide, including a functional fragment or avariant thereof, or a membrane domain, including a functional fragmentor a variant thereof, encoded by a reference nucleotide sequence that isnot sequence optimized), e.g., improved properties related to expressionefficacy after administration in vivo. Such properties include, but arenot limited to, improving nucleic acid stability (e.g., mRNA stability),increasing translation efficacy in the target tissue, reducing thenumber of truncated proteins expressed, improving the folding or preventmisfolding of the expressed proteins, reducing toxicity of the expressedproducts, reducing cell death caused by the expressed products,increasing and/or decreasing protein aggregation.

In some embodiments, the sequence-optimized nucleotide sequence is codonoptimized for expression in human subjects, having structural and/orchemical features that avoid one or more of the problems in the art, forexample, features which are useful for optimizing formulation anddelivery of nucleic acid-based therapeutics while retaining structuraland functional integrity; overcoming a threshold of expression;improving expression rates; half-life and/or protein concentrations;optimizing protein localization; and avoiding deleterious bio-responsessuch as the immune response and/or degradation pathways.

In some embodiments, the polynucleotides of the disclosure comprise anucleotide sequence (e.g., a nucleotide sequence encoding an IL-12polypeptide, a nucleotide sequence encoding a membrane domain, anucleotide sequence encoding an additional polypeptide of interest, a5′-UTR, a 3′-UTR, a microRNA, a nucleic acid sequence encoding a linker,or any combination thereof) that is sequence-optimized according to amethod comprising:

(i) substituting at least one codon in a reference nucleotide sequence(e.g., a nucleic acid sequence encoding an IL-12 polypeptide and/or anucleic acid sequence encoding a membrane domain) with an alternativecodon to increase or decrease uridine content to generate auridine-modified sequence;

(ii) substituting at least one codon in a reference nucleotide sequence(e.g., a nucleic acid sequence encoding an IL-12 polypeptide and/or anucleic acid sequence encoding a membrane domain) with an alternativecodon having a higher codon frequency in the synonymous codon set;

(iii) substituting at least one codon in a reference nucleotide sequence(e.g., a nucleic acid sequence encoding an IL-12 polypeptide and/or anucleic acid sequence encoding a membrane domain) with an alternativecodon to increase G/C content; or

(iv) a combination thereof.

In some embodiments, the sequence-optimized nucleotide sequence (e.g., anucleic acid sequence encoding an IL-12 polypeptide and/or a nucleicacid sequence encoding a membrane domain) has at least one improvedproperty with respect to the reference nucleotide sequence.

In some embodiments, the sequence optimization method is multiparametricand comprises one, two, three, four, or more methods disclosed hereinand/or other optimization methods known in the art.

Features, which can be considered beneficial in some embodiments of thedisclosure, can be encoded by or within regions of the polynucleotideand such regions can be upstream (5′) to, downstream (3′) to, or withinthe region that encodes the IL-12 polypeptide and/or the region thatencodes membrane domain. These regions can be incorporated into thepolynucleotide before and/or after sequence-optimization of the proteinencoding region or open reading frame (ORF). Examples of such featuresinclude, but are not limited to, untranslated regions (UTRs), microRNAsequences, Kozak sequences, oligo(dT) sequences, poly-A tail, anddetectable tags and can include multiple cloning sites that may haveXbaI recognition.

In some embodiments, the polynucleotide of the disclosure comprises a 5′UTR, a 3′ UTR and/or a miRNA. In some embodiments, the polynucleotidecomprises two or more 5′ UTRs and/or 3′ UTRs, which can be the same ordifferent sequences. In some embodiments, the polynucleotide comprisestwo or more miRNA, which can be the same or different sequences. Anyportion of the 5′ UTR, 3′ UTR, and/or miRNA, including none, can besequence-optimized and can independently contain one or more differentstructural or chemical modifications, before and/or after sequenceoptimization.

In some embodiments, after optimization, the polynucleotide isreconstituted and transformed into a vector such as, but not limited to,plasmids, viruses, cosmids, and artificial chromosomes. For example, theoptimized polynucleotide can be reconstituted and transformed intochemically competent E. coli, yeast, neurospora, maize, drosophila, etc.where high copy plasmid-like or chromosome structures occur by methodsdescribed herein.

9. Sequence-Optimized Nucleotide Sequences Encoding IL-12 Polypeptides

In some embodiments, the polynucleotide of the disclosure comprises asequence-optimized nucleotide sequence encoding an IL-12 polypeptide(i.e., an IL-12B and/or IL-12A polypeptide) disclosed herein. In someembodiments, the polynucleotide of the disclosure comprises a nucleicacid sequence encoding an IL-12B and/or a nucleic acid sequence encodingan IL-12A polypeptide and a nucleic acid sequence encoding a membranedomain, wherein the nucleic acid sequences have been sequence optimized.

Exemplary sequence-optimized nucleotide sequences encoding human IL-12Band/or IL-12A are set forth in SEQ ID NOs: 5-44, 98-100, 104-180, 220and 221. In some embodiments, the sequence optimized IL-12B and/orIL-12A sequences set forth in SEQ ID NOs: 5-44, 98-100, 104-180, 220 and221, fragments, and variants thereof are used to practice the methodsdisclosed herein. In some embodiments, the sequence optimized IL-12Band/or IL-12A sequences set forth in SEQ ID NOs: 5-44, 98-100, 104-180,220 and 221, fragments and variants thereof are combined with oralternatives to the wild-type sequences disclosed in SEQ ID NOs: 1-6.Based on the RNA sequences provided, a person of ordinary skill in theart would understand the corresponding DNA sequence (e.g., conversion ofuracil to thymine). Likewise, based on the DNA sequences provided, aperson of ordinary skill in the art would understand the correspondingRNA sequence (e.g., conversion of thymine to uracil).

The sequence-optimized nucleotide sequences disclosed herein aredistinct from the corresponding wild type nucleotide acid sequences andfrom other known sequence-optimized nucleotide sequences, e.g., thesesequence-optimized nucleic acids have unique compositionalcharacteristics.

10. Methods for Sequence Optimization

In some embodiments, a polynucleotide of the disclosure (e.g., apolynucleotide comprising a nucleotide sequence encoding an IL-12polypeptide comprising IL-12A, IL-12B, or both IL-12A and IL-12B (e.g.,the wild-type sequence, functional fragment, or variant thereof) and amembrane domain) is sequence optimized. A sequence optimized nucleotidesequence (nucleotide sequence is also referred to as “nucleic acid”herein) comprises at least one codon modification with respect to areference sequence (e.g., a wild-type sequence encoding an IL-12polypeptide comprising IL-12A, IL-12B, or both IL-12A and IL-12B or amembrane domain). Thus, in a sequence optimized nucleic acid, at leastone codon is different from a corresponding codon in a referencesequence (e.g., a wild-type sequence).

In general, sequence optimized nucleic acids are generated by at least astep comprising substituting codons in a reference sequence withsynonymous codons (i.e., codons that encode the same amino acid). Suchsubstitutions can be effected, for example, by applying a codonsubstitution map (i.e., a table providing the codons that will encodeeach amino acid in the codon optimized sequence), or by applying a setof rules (e.g., if glycine is next to neutral amino acid, glycine wouldbe encoded by a certain codon, but if it is next to a polar amino acid,it would be encoded by another codon). In addition to codonsubstitutions (i.e., “codon optimization”) the sequence optimizationmethods disclosed herein comprise additional optimization steps whichare not strictly directed to codon optimization such as the removal ofdeleterious motifs (destabilizing motif substitution). Compositions andformulations comprising these sequence optimized nucleic acids (e.g., aRNA, e.g., an mRNA) can be administered to a subject in need thereof tofacilitate in vivo expression of functionally active IL-12.

The recombinant expression of large molecules in cell cultures can be achallenging task with numerous limitations (e.g., poor proteinexpression levels, stalled translation resulting in truncated expressionproducts, protein misfolding, etc.) These limitations can be reduced oravoided by administering the polynucleotides (e.g., a RNA, e.g., anmRNA), which encode a functionally active IL-12 or compositions orformulations comprising the same to a patient suffering from cancer, sothe synthesis and delivery of the IL-12 polypeptide to treat cancertakes place endogenously.

Changing from an in vitro expression system (e.g., cell culture) to invivo expression requires the redesign of the nucleic acid sequenceencoding the therapeutic agent. Redesigning a naturally occurring genesequence by choosing different codons without necessarily altering theencoded amino acid sequence can often lead to dramatic increases inprotein expression levels (Gustafsson et al., 2004, Journal/TrendsBiotechnol 22, 346-53). Variables such as codon adaptation index (CAI),mRNA secondary structures, cis-regulatory sequences, GC content and manyother similar variables have been shown to somewhat correlate withprotein expression levels (Villalobos et al., 2006, “Journal/BMCBioinformatics 7, 285). However, due to the degeneracy of the geneticcode, there are numerous different nucleic acid sequences that can allencode the same therapeutic agent. Each amino acid is encoded by up tosix synonymous codons; and the choice between these codons influencesgene expression. In addition, codon usage (i.e., the frequency withwhich different organisms use codons for expressing a polypeptidesequence) differs among organisms (for example, recombinant productionof human or humanized therapeutic antibodies frequently takes place inhamster cell cultures).

In some embodiments, a reference nucleic acid sequence can be sequenceoptimized by applying a codon map. The skilled artisan will appreciatethat the T bases in the codon maps disclosed below are present in DNA,whereas the T bases would be replaced by U bases in corresponding RNAs.For example, a sequence optimized nucleic acid disclosed herein in DNAform, e.g., a vector or an in-vitro translation (IVT) template, wouldhave its T bases transcribed as U based in its corresponding transcribedmRNA. In this respect, both sequence optimized DNA sequences (comprisingT) and their corresponding RNA sequences (comprising U) are consideredsequence optimized nucleic acid of the present disclosure. A skilledartisan would also understand that equivalent codon-maps can begenerated by replaced one or more bases with non-natural bases. Thus,e.g., a TTC codon (DNA map) would correspond to a UUC codon (RNA map),which in turn may correspond to a ΨΨC codon (RNA map in which U has beenreplaced with pseudouridine).

In one embodiment, a reference sequence encoding IL-12A, IL-12B, or bothIL-12A and IL-12B and/or a membrane domain can be optimized by replacingall the codons encoding a certain amino acid with only one of thealternative codons provided in a codon map. For example, all the valinesin the optimized sequence would be encoded by GTG or GTC or GTT.

Sequence optimized polynucleotides of the disclosure can be generatedusing one or more codon optimization methods, or a combination thereof.Sequence optimization methods which may be used to sequence optimizenucleic acid sequences are described in detail herein. This list ofmethods is not comprehensive or limiting.

It will be appreciated that the design principles and rules describedfor each one of the sequence optimization methods discussed below can becombined in many different ways, for example high G/C content sequenceoptimization for some regions or uridine content sequence optimizationfor other regions of the reference nucleic acid sequence, as well astargeted nucleotide mutations to minimize secondary structure throughoutthe sequence or to eliminate deleterious motifs.

The choice of potential combinations of sequence optimization methodscan be, for example, dependent on the specific chemistry used to producea synthetic polynucleotide. Such a choice can also depend oncharacteristics of the protein encoded by the sequence optimized nucleicacid, e.g., a full sequence, a functional fragment, or a fusion proteincomprising IL-12, etc. In some embodiments, such a choice can depend onthe specific tissue or cell targeted by the sequence optimized nucleicacid (e.g., a therapeutic synthetic mRNA).

The mechanisms of combining the sequence optimization methods or designrules derived from the application and analysis of the optimizationmethods can be either simple or complex. For example, the combinationcan be:

-   -   (i) Sequential: Each sequence optimization method or set of        design rules applies to a different subsequence of the overall        sequence, for example reducing uridine at codon positions 1 to        30 and then selecting high frequency codons for the remainder of        the sequence;    -   (ii) Hierarchical: Several sequence optimization methods or sets        of design rules are combined in a hierarchical, deterministic        fashion. For example, use the most GC-rich codons, breaking ties        (which are common) by choosing the most frequent of those        codons.    -   (iii) Multifactorial/Multiparametric: Machine learning or other        modeling techniques are used to design a single sequence that        best satisfies multiple overlapping and possibly contradictory        requirements. This approach would require the use of a computer        applying a number of mathematical techniques, for example,        genetic algorithms.

Ultimately, each one of these approaches can result in a specific set ofrules which in many cases can be summarized in a single codon table,i.e., a sorted list of codons for each amino acid in the target protein(i.e., an IL-12A polypeptide, an IL-12B polypeptide, both IL-12A andIL-12B polypeptides, and/or a membrane domain), with a specific rule orset of rules indicating how to select a specific codon for each aminoacid position.

a. Codon Frequency—Codon Usage Bias

Numerous codon optimization methods known in the art are based on thesubstitution of codons in a reference nucleic acid sequence with codonshaving higher frequencies. Thus, in some embodiments, a nucleic acidsequence disclosed herein (e.g., a nucleic acid sequence encoding anIL-12 polypeptide and/or a nucleic acid sequence encoding a membranedomain) can be sequence optimized using methods comprising the use ofmodifications in the frequency of use of one or more codons relative toother synonymous codons in the sequence optimized nucleic acid withrespect to the frequency of use in the non-codon optimized sequence.

As used herein, the term “codon frequency” refers to codon usage bias,i.e., the differences in the frequency of occurrence of synonymouscodons in coding DNA/RNA. It is generally acknowledged that codonpreferences reflect a balance between mutational biases and naturalselection for translational optimization. Optimal codons help to achievefaster translation rates and high accuracy. As a result of thesefactors, translational selection is expected to be stronger in highlyexpressed genes. In the field of bioinformatics and computationalbiology, many statistical methods have been proposed and used to analyzecodon usage bias. See, e.g., Comeron & Aguadé (1998) J. Mol. Evol. 47:268-74. Methods such as the ‘frequency of optimal codons’ (Fop) (Ikemura(1981) J. Mol. Biol. 151 (3): 389-409), the Relative Codon Adaptation(RCA) (Fox & Eril (2010) DNA Res. 17 (3): 185-96) or the ‘CodonAdaptation Index’ (CAI) (Sharp & Li (1987) Nucleic Acids Res. 15 (3):1281-95) are used to predict gene expression levels, while methods suchas the ‘effective number of codons’ (Nc) and Shannon entropy frominformation theory are used to measure codon usage evenness.Multivariate statistical methods, such as correspondence analysis andprincipal component analysis, are widely used to analyze variations incodon usage among genes (Suzuki et al. (2008) DNA Res. 15 (6): 357-65;Sandhu et al., In Silico Biol. 2008; 8(2):187-92).

A nucleic acid sequence disclosed herein (e.g., a wild type nucleic acidsequence, a mutant nucleic acid sequence, a chimeric nucleic sequence,etc. which can be, for example, an mRNA), can be codon optimized usingmethods comprising substituting at least one codon in the referencenucleic acid sequence with an alternative codon having a higher or lowercodon frequency in the synonymous codon set; wherein the resultingsequence optimized nucleic acid has at least one optimized property withrespect to the reference nucleic acid sequence.

In some embodiments, at least about 5%, at least about 10%, at leastabout 15%, at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at leastabout 50%, at least about 55%, at least about 60%, at least about 65%,at least about 70%, at least about 75%, at least about 80%, at leastabout 85%, at least about 90%, at least about 95%, at least about 99%,or 100% of the codons in the reference nucleic acid sequence aresubstituted with alternative codons, each alternative codon having acodon frequency higher than the codon frequency of the substituted codonin the synonymous codon set.

In some embodiments, at least one codon in the reference nucleic acidsequence is substituted with an alternative codon having a codonfrequency higher than the codon frequency of the substituted codon inthe synonymous codon set, and at least one codon in the referencenucleic acid sequence is substituted with an alternative codon having acodon frequency lower than the codon frequency of the substituted codonin the synonymous codon set.

In some embodiments, at least about 5%, at least about 10%, at leastabout 15%, at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at leastabout 50%, at least about 55%, at least about 60%, at least about 65%,at least about 70%, or at least about 75% of the codons in the referencenucleic acid sequence are substituted with alternative codons, eachalternative codon having a codon frequency higher than the codonfrequency of the substituted codon in the synonymous codon set.

In some embodiments, at least one alternative codon having a highercodon frequency has the highest codon frequency in the synonymous codonset. In other embodiments, all alternative codons having a higher codonfrequency have the highest codon frequency in the synonymous codon set.

In some embodiments, at least one alternative codon having a lower codonfrequency has the lowest codon frequency in the synonymous codon set. Insome embodiments, all alternative codons having a higher codon frequencyhave the highest codon frequency in the synonymous codon set.

In some specific embodiments, at least one alternative codon has thesecond highest, the third highest, the fourth highest, the fifth highestor the sixth highest frequency in the synonymous codon set. In somespecific embodiments, at least one alternative codon has the secondlowest, the third lowest, the fourth lowest, the fifth lowest, or thesixth lowest frequency in the synonymous codon set.

Optimization based on codon frequency can be applied globally, asdescribed above, or locally to the reference nucleic acid sequence. Insome embodiments, when applied locally, regions of the reference nucleicacid sequence can modified based on codon frequency, substituting all ora certain percentage of codons in a certain subsequence with codons thathave higher or lower frequencies in their respective synonymous codonsets. Thus, in some embodiments, at least about 5%, at least about 10%,at least about 15%, at least about 20%, at least about 25%, at leastabout 30%, at least about 35%, at least about 40%, at least about 45%,at least about 50%, at least about 55%, at least about 60%, at leastabout 65%, at least about 70%, at least about 75%, at least about 80%,at least about 85%, at least about 90%, at least about 95%, at leastabout 99%, or 100% of the codons in a subsequence of the referencenucleic acid sequence are substituted with alternative codons, eachalternative codon having a codon frequency higher than the codonfrequency of the substituted codon in the synonymous codon set.

In some embodiments, at least one codon in a subsequence of thereference nucleic acid sequence is substituted with an alternative codonhaving a codon frequency higher than the codon frequency of thesubstituted codon in the synonymous codon set, and at least one codon ina subsequence of the reference nucleic acid sequence is substituted withan alternative codon having a codon frequency lower than the codonfrequency of the substituted codon in the synonymous codon set.

In some embodiments, at least about 5%, at least about 10%, at leastabout 15%, at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at leastabout 50%, at least about 55%, at least about 60%, at least about 65%,at least about 70%, or at least about 75% of the codons in a subsequenceof the reference nucleic acid sequence are substituted with alternativecodons, each alternative codon having a codon frequency higher than thecodon frequency of the substituted codon in the synonymous codon set.

In some embodiments, at least one alternative codon substituted in asubsequence of the reference nucleic acid sequence and having a highercodon frequency has the highest codon frequency in the synonymous codonset. In other embodiments, all alternative codons substituted in asubsequence of the reference nucleic acid sequence and having a lowercodon frequency have the lowest codon frequency in the synonymous codonset.

In some embodiments, at least one alternative codon substituted in asubsequence of the reference nucleic acid sequence and having a lowercodon frequency has the lowest codon frequency in the synonymous codonset. In some embodiments, all alternative codons substituted in asubsequence of the reference nucleic acid sequence and having a highercodon frequency have the highest codon frequency in the synonymous codonset.

In specific embodiments, a sequence optimized nucleic acid can comprisea subsequence having an overall codon frequency higher or lower than theoverall codon frequency in the corresponding subsequence of thereference nucleic acid sequence at a specific location, for example, atthe 5′ end or 3′ end of the sequence optimized nucleic acid, or within apredetermined distance from those region (e.g., at least 1, 2, 3, 4, 5,6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80,85, 90, 95 or 100 codons from the 5′ end or 3′ end of the sequenceoptimized nucleic acid).

In some embodiments, a sequence optimized nucleic acid can comprise morethan one subsequence having an overall codon frequency higher or lowerthan the overall codon frequency in the corresponding subsequence of thereference nucleic acid sequence. A skilled artisan would understand thatsubsequences with overall higher or lower overall codon frequencies canbe organized in innumerable patterns, depending on whether the overallcodon frequency is higher or lower, the length of the subsequence, thedistance between subsequences, the location of the subsequences, etc.

See, U.S. Pat. Nos. 5,082,767, 8,126,653, 7,561,973, 8,401,798; U.S.Publ. No. US 20080046192, US 20080076161; Int'l. Publ. No. WO2000018778;Welch et al. (2009) PLoS ONE 4(9): e7002; Gustafsson et al. (2012)Protein Expression and Purification 83: 37-46; Chung et al. (2012) BMCSystems Biology 6:134; all of which are incorporated herein by referencein their entireties.

b. Destabilizing Motif Substitution

There is a variety of motifs that can affect sequence optimization,which fall into various non-exclusive categories, for example:

-   -   (i) Primary sequence based motifs: Motifs defined by a simple        arrangement of nucleotides.    -   (ii) Structural motifs: Motifs encoded by an arrangement of        nucleotides that tends to form a certain secondary structure.    -   (iii) Local motifs: Motifs encoded in one contiguous        subsequence.    -   (iv) Distributed motifs: Motifs encoded in two or more disjoint        subsequences.    -   (v) Advantageous motifs: Motifs which improve nucleotide        structure or function.    -   (vi) Disadvantageous motifs: Motifs with detrimental effects on        nucleotide structure or function.

There are many motifs that fit into the category of disadvantageousmotifs. Some examples include, for example, restriction enzyme motifs,which tend to be relatively short, exact sequences such as therestriction site motifs for Xba1 (TCTAGA (SEQ ID NO: 187)), EcoRI(GAATTC (SEQ ID NO: 188)), EcoRII (CCWGG (SEQ ID NO: 189), wherein Wmeans A or T, per the IUPAC ambiguity codes), or HindIII (AAGCTT (SEQ IDNO: 190)); enzyme sites, which tend to be longer and based on consensusnot exact sequence, such in the T7 RNA polymerase (GnnnnWnCRnCTCnCnnWnD(SEQ ID NO: 191), wherein n means any nucleotide, R means A or G, Wmeans A or T, D means A or G or T but not C); structural motifs, such asGGGG repeats (Kim et al. (1991) Nature 351(6324):331-2); or other motifssuch as CUG-triplet repeats (Querido et al. (2014) J. Cell Sci.124:1703-1714).

Accordingly, a nucleic acid sequence disclosed herein can be sequenceoptimized using methods comprising substituting at least onedestabilizing motif in a reference nucleic acid sequence, and removingsuch disadvantageous motif or replacing it with an advantageous motif.

In some embodiments, the optimization process comprises identifyingadvantageous and/or disadvantageous motifs in the reference nucleicsequence, wherein such motifs are, e.g., specific subsequences that cancause a loss of stability in the reference nucleic acid sequence prioror during the optimization process. For example, substitution ofspecific bases during optimization may generate a subsequence (motif)recognized by a restriction enzyme. Accordingly, during the optimizationprocess the appearance of disadvantageous motifs can be monitored bycomparing the sequence optimized sequence with a library of motifs knownto be disadvantageous. Then, the identification of disadvantageousmotifs could be used as a post-hoc filter, i.e., to determine whether acertain modification which potentially could be introduced in thereference nucleic acid sequence should be actually implemented or not.

In some embodiments, the identification of disadvantageous motifs can beused prior to the application of the sequence optimization methodsdisclosed herein, i.e., the identification of motifs in the referencenucleic acid sequence and their replacement with alternative nucleicacid sequences can be used as a preprocessing step, for example, beforeuridine reduction.

In other embodiments, the identification of disadvantageous motifs andtheir removal is used as an additional sequence optimization techniqueintegrated in a multiparametric nucleic acid optimization methodcomprising two or more of the sequence optimization methods disclosedherein. When used in this fashion, a disadvantageous motif identifiedduring the optimization process would be removed, for example, bysubstituting the lowest possible number of nucleobases in order topreserve as closely as possible the original design principle(s) (e.g.,low U, high frequency, etc.).

See, e.g., U.S. Publ. Nos. US20140228558, US20050032730, orUS20140228558, which are herein incorporated by reference in theirentireties.

c. Limited Codon Set Optimization

In some particular embodiments, sequence optimization of a referencenucleic acid sequence can be conducted using a limited codon set, e.g.,a codon set wherein less than the native number of codons is used toencode the 20 natural amino acids, a subset of the 20 natural aminoacids, or an expanded set of amino acids including, for example,non-natural amino acids.

The genetic code is highly similar among all organisms and can beexpressed in a simple table with 64 entries which would encode the 20standard amino acids involved in protein translation plus start and stopcodons. The genetic code is degenerate, i.e., in general, more than onecodon specifies each amino acid. For example, the amino acid leucine isspecified by the UUA, UUG, CUU, CUC, CUA, or CUG codons, while the aminoacid serine is specified by UCA, UCG, UCC, UCU, AGU, or AGC codons(difference in the first, second, or third position). Native geneticcodes comprise 62 codons encoding naturally occurring amino acids. Thus,in some embodiments of the methods disclosed herein optimized codon sets(genetic codes) comprising less than 62 codons to encode 20 amino acidscan comprise 61, 60, 59, 58, 57, 56, 55, 54, 53, 52, 51, 50, 49, 48, 47,46, 45, 44, 43, 42, 41, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 30,29, 28, 27, 26, 25, 24, 23, 22, 21, or 20 codons.

In some embodiments, the limited codon set comprises less than 20codons. For example, if a protein contains less than 20 types of aminoacids, such protein could be encoded by a codon set with less than 20codons. Accordingly, in some embodiments, an optimized codon setcomprises as many codons as different types of amino acids are presentin the protein encoded by the reference nucleic acid sequence. In someembodiments, the optimized codon set comprises 19, 18, 17, 16, 15, 14,13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or even 1 codon.

In some embodiments, at least one amino acid selected from the groupconsisting of Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu,Lys, Phe, Pro, Ser, Thr, Tyr, and Val, i.e., amino acids which arenaturally encoded by more than one codon, is encoded with less codonsthan the naturally occurring number of synonymous codons. For example,in some embodiments, Ala can be encoded in the sequence optimizednucleic acid by 3, 2 or 1 codons; Cys can be encoded in the sequenceoptimized nucleic acid by 1 codon; Asp can be encoded in the sequenceoptimized nucleic acid by 1 codon; Glu can be encoded in the sequenceoptimized nucleic acid by 1 codon; Phe can be encoded in the sequenceoptimized nucleic acid by 1 codon; Gly can be encoded in the sequenceoptimized nucleic acid by 3 codons, 2 codons or 1 codon; His can beencoded in the sequence optimized nucleic acid by 1 codon; Ile can beencoded in the sequence optimized nucleic acid by 2 codons or 1 codon;Lys can be encoded in the sequence optimized nucleic acid by 1 codon;Leu can be encoded in the sequence optimized nucleic acid by 5 codons, 4codons, 3 codons, 2 codons or 1 codon; Asn can be encoded in thesequence optimized nucleic acid by 1 codon; Pro can be encoded in thesequence optimized nucleic acid by 3 codons, 2 codons, or 1 codon; Glncan be encoded in the sequence optimized nucleic acid by 1 codon; Argcan be encoded in the sequence optimized nucleic acid by 5 codons, 4codons, 3 codons, 2 codons, or 1 codon; Ser can be encoded in thesequence optimized nucleic acid by 5 codons, 4 codons, 3 codons, 2codons, or 1 codon; Thr can be encoded in the sequence optimized nucleicacid by 3 codons, 2 codons, or 1 codon; Val can be encoded in thesequence optimized nucleic acid by 3 codons, 2 codons, or 1 codon; and,Tyr can be encoded in the sequence optimized nucleic acid by 1 codon.

In some embodiments, at least one amino acid selected from the groupconsisting of Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu,Lys, Phe, Pro, Ser, Thr, Tyr, and Val, i.e., amino acids which arenaturally encoded by more than one codon, is encoded by a single codonin the limited codon set.

In some specific embodiments, the sequence optimized nucleic acid is aDNA and the limited codon set consists of 20 codons, wherein each codonencodes one of 20 amino acids. In some embodiments, the sequenceoptimized nucleic acid is a DNA and the limited codon set comprises atleast one codon selected from the group consisting of GCT, GCC, GCA, andGCG; at least a codon selected from the group consisting of CGT, CGC,CGA, CGG, AGA, and AGG; at least a codon selected from AAT or ACC; atleast a codon selected from GAT or GAC; at least a codon selected fromTGT or TGC; at least a codon selected from CAA or CAG; at least a codonselected from GAA or GAG; at least a codon selected from the groupconsisting of GGT, GGC, GGA, and GGG; at least a codon selected from CATor CAC; at least a codon selected from the group consisting of ATT, ATC,and ATA; at least a codon selected from the group consisting of TTA,TTG, CTT, CTC, CTA, and CTG; at least a codon selected from AAA or AAG;an ATG codon; at least a codon selected from TTT or TTC; at least acodon selected from the group consisting of CCT, CCC, CCA, and CCG; atleast a codon selected from the group consisting of TCT, TCC, TCA, TCG,AGT, and AGC; at least a codon selected from the group consisting ofACT, ACC, ACA, and ACG; a TGG codon; at least a codon selected from TATor TAC; and, at least a codon selected from the group consisting of GTT,GTC, GTA, and GTG.

In other embodiments, the sequence optimized nucleic acid is an RNA(e.g., an mRNA) and the limited codon set consists of 20 codons, whereineach codon encodes one of 20 amino acids. In some embodiments, thesequence optimized nucleic acid is an RNA and the limited codon setcomprises at least one codon selected from the group consisting of GCU,GCC, GCA, and GCG; at least a codon selected from the group consistingof CGU, CGC, CGA, CGG, AGA, and AGG; at least a codon selected from AAUor ACC; at least a codon selected from GAU or GAC; at least a codonselected from UGU or UGC; at least a codon selected from CAA or CAG; atleast a codon selected from GAA or GAG; at least a codon selected fromthe group consisting of GGU, GGC, GGA, and GGG; at least a codonselected from CAU or CAC; at least a codon selected from the groupconsisting of AUU, AUC, and AUA; at least a codon selected from thegroup consisting of UUA, UUG, CUU, CUC, CUA, and CUG; at least a codonselected from AAA or AAG; an AUG codon; at least a codon selected fromUUU or UUC; at least a codon selected from the group consisting of CCU,CCC, CCA, and CCG; at least a codon selected from the group consistingof UCU, UCC, UCA, UCG, AGU, and AGC; at least a codon selected from thegroup consisting of ACU, ACC, ACA, and ACG; a UGG codon; at least acodon selected from UAU or UAC; and, at least a codon selected from thegroup consisting of GUU, GUC, GUA, and GUG.

In some specific embodiments, the limited codon set has been optimizedfor in vivo expression of a sequence optimized nucleic acid (e.g., asynthetic mRNA) following administration to a certain tissue or cell.

In some embodiments, the optimized codon set (e.g., a 20 codon setencoding 20 amino acids) complies at least with one of the followingproperties:

-   -   (a) the optimized codon set has a higher average G/C content        than the original or native codon set; or,    -   (b) the optimized codon set has a lower average U content than        the original or native codon set; or,    -   (c) the optimized codon set is composed of codons with the        highest frequency; or,    -   (d) the optimized codon set is composed of codons with the        lowest frequency; or,    -   (e) a combination thereof.

In some specific embodiments, at least one codon in the optimized codonset has the second highest, the third highest, the fourth highest, thefifth highest or the sixth highest frequency in the synonymous codonset. In some specific embodiments, at least one codon in the optimizedcodon has the second lowest, the third lowest, the fourth lowest, thefifth lowest, or the sixth lowest frequency in the synonymous codon set.

As used herein, the term “native codon set” refers to the codon set usednatively by the source organism to encode the reference nucleic acidsequence. As used herein, the term “original codon set” refers to thecodon set used to encode the reference nucleic acid sequence before thebeginning of sequence optimization, or to a codon set used to encode anoptimized variant of the reference nucleic acid sequence at thebeginning of a new optimization iteration when sequence optimization isapplied iteratively or recursively.

In some embodiments, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%,55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% of codons in thecodon set are those with the highest frequency. In other embodiments,5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%,75%, 80%, 85%, 90%, 95% or 100% of codons in the codon set are thosewith the lowest frequency.

In some embodiments, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%,55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% of codons in thecodon set are those with the highest uridine content. In someembodiments, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%,65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% of codons in the codon set arethose with the lowest uridine content.

In some embodiments, the average G/C content (absolute or relative) ofthe codon set is 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%,60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% higher than the averageG/C content (absolute or relative) of the original codon set. In someembodiments, the average G/C content (absolute or relative) of the codonset is 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%,70%, 75%, 80%, 85%, 90%, 95% or 100% lower than the average G/C content(absolute or relative) of the original codon set.

In some embodiments, the uracil content (absolute or relative) of thecodon set is 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%,65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% higher than the average uracilcontent (absolute or relative) of the original codon set. In someembodiments, the uracil content (absolute or relative) of the codon setis 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%,75%, 80%, 85%, 90%, 95% or 100% lower than the average uracil content(absolute or relative) of the original codon set.

See also U.S. Appl. Publ. No. 2011/0082055, and Int'l. Publ. No.WO2000018778, both of which are incorporated herein by reference intheir entireties.

11. Characterization of Sequence Optimized Nucleic Acids

In some embodiments of the disclosure, the polynucleotide (e.g., a RNA,e.g., an mRNA) comprising a sequence optimized nucleic acid disclosedherein can be can be tested to determine whether at least one nucleicacid sequence property (e.g., stability when exposed to nucleases) orexpression property has been improved with respect to the non-sequenceoptimized nucleic acid.

As used herein, “expression property” refers to a property of a nucleicacid sequence either in vivo (e.g., translation efficacy of a syntheticmRNA after administration to a subject in need thereof) or in vitro(e.g., translation efficacy of a synthetic mRNA tested in an in vitromodel system). Expression properties include but are not limited to theamount of protein produced by an mRNA after administration, and theamount of soluble or otherwise functional protein produced. In someembodiments, sequence optimized nucleic acids disclosed herein can beevaluated according to the viability of the cells expressing a proteinencoded by a sequence optimized nucleic acid sequence (e.g., a RNA,e.g., an mRNA) disclosed herein.

In a particular embodiment, a plurality of sequence optimized nucleicacids disclosed herein (e.g., a RNA, e.g., an mRNA) containing codonsubstitutions with respect to the non-optimized reference nucleic acidsequence can be characterized functionally to measure a property ofinterest, for example an expression property in an in vitro modelsystem, or in vivo in a target tissue or cell.

a. Optimization of Nucleic Acid Sequence Intrinsic Properties

In some embodiments of the disclosure, the desired property of thepolynucleotide is an intrinsic property of the nucleic acid sequence.For example, the nucleotide sequence (e.g., a RNA, e.g., an mRNA) can besequence optimized for in vivo or in vitro stability. In someembodiments, the nucleotide sequence can be sequence optimized forexpression in a particular target tissue or cell. In some embodiments,the nucleic acid sequence is sequence optimized to increase its plasmahalf by preventing its degradation by endo and exonucleases.

In other embodiments, the nucleic acid sequence is sequence optimized toincrease its resistance to hydrolysis in solution, for example, tolengthen the time that the sequence optimized nucleic acid or apharmaceutical composition comprising the sequence optimized nucleicacid can be stored under aqueous conditions with minimal degradation.

In other embodiments, the sequence optimized nucleic acid can beoptimized to increase its resistance to hydrolysis in dry storageconditions, for example, to lengthen the time that the sequenceoptimized nucleic acid can be stored after lyophilization with minimaldegradation.

b. Nucleic Acids Sequence Optimized for Protein Expression

In some embodiments of the disclosure, the desired property of thepolynucleotide is the level of expression of an IL-12A polypeptide, anIL-12B polypeptide, or both IL-12A and IL-12B polypeptides and amembrane domain encoded by a sequence optimized sequence disclosedherein. Protein expression levels can be measured using one or moreexpression systems. In some embodiments, expression can be measured incell culture systems, e.g., CHO cells or HEK293 cells. In someembodiments, expression can be measured using in vitro expressionsystems prepared from extracts of living cells, e.g., rabbitreticulocyte lysates, or in vitro expression systems prepared byassembly of purified individual components. In other embodiments, theprotein expression is measured in an in vivo system, e.g., mouse,rabbit, monkey, etc.

In some embodiments, protein expression in solution form can bedesirable. Accordingly, in some embodiments, a reference sequence can besequence optimized to yield a sequence optimized nucleic acid sequencehaving optimized levels of expressed proteins in soluble form. Levels ofprotein expression and other properties such as solubility, levels ofaggregation, and the presence of truncation products (i.e., fragmentsdue to proteolysis, hydrolysis, or defective translation) can bemeasured according to methods known in the art, for example, usingelectrophoresis (e.g., native or SDS-PAGE) or chromatographic methods(e.g., HPLC, size exclusion chromatography, etc.).

c. Optimization of Target Tissue or Target Cell Viability

In some embodiments, the expression of heterologous therapeutic proteinsencoded by a nucleic acid sequence can have deleterious effects in thetarget tissue or cell, reducing protein yield, or reducing the qualityof the expressed product (e.g., due to the presence of protein fragmentsor precipitation of the expressed protein in inclusion bodies), orcausing toxicity.

Accordingly, in some embodiments of the disclosure, the sequenceoptimization of a nucleic acid sequence disclosed herein, e.g., anucleic acid sequence encoding an IL-12 polypeptide and/or a nucleicacid sequence encoding a membrane domain, can be used to increase theviability of target cells expressing the protein encoded by the sequenceoptimized nucleic acid.

Heterologous protein expression can also be deleterious to cellstransfected with a nucleic acid sequence for autologous or heterologoustransplantation. Accordingly, in some embodiments of the presentdisclosure the sequence optimization of a nucleic acid sequencedisclosed herein can be used to increase the viability of target cellsexpressing the protein encoded by the sequence optimized nucleic acidsequence. Changes in cell or tissue viability, toxicity, and otherphysiological reaction can be measured according to methods known in theart.

d. Reduction of Immune and/or Inflammatory Response

In some cases, the administration of a sequence optimized nucleic acidencoding a tethered IL-12 polypeptide as disclosed herein, includingfunctional fragments and variants thereof, may trigger an immuneresponse, which could be caused by (i) the therapeutic agent (e.g., anmRNA comprising a nucleic acid sequence encoding a tethered IL-12polypeptide), or (ii) the expression product of such therapeutic agent(e.g., the tethered IL-12 polypeptide encoded by the mRNA), or (iv) acombination thereof. Accordingly, in some embodiments of the presentdisclosure the sequence optimization of nucleic acid sequence (e.g., anmRNA) disclosed herein can be used to decrease an immune or inflammatoryresponse triggered by the administration of a nucleic acid encoding atethered IL-12 polypeptide or by the expression product of the tetheredIL-12 polypeptide encoded by such nucleic acid.

In some aspects, an inflammatory response can be measured by detectingincreased levels of one or more inflammatory cytokines using methodsknown in the art, e.g., ELISA. The term “inflammatory cytokine” refersto cytokines that are elevated in an inflammatory response. Examples ofinflammatory cytokines include interleukin-6 (IL-6), CXCL1 (chemokine(C-X-C motif) ligand 1; also known as GROα, interferon-γ (IFNγ), tumornecrosis factor α (TNFα), interferon γ-induced protein 10 (IP-10), orgranulocyte-colony stimulating factor (G-CSF). The term inflammatorycytokines includes also other cytokines associated with inflammatoryresponses known in the art, e.g., interleukin-1 (IL-1), interleukin-8(IL-8), interleukin-13 (11-13), interferon α (IFN-α), etc.

12. Methods for Modifying Polynucleotides

The disclosure includes modified polynucleotides comprising apolynucleotide described herein (e.g., a polynucleotide comprising anucleotide sequence encoding an IL-12 polypeptide and a nucleotidesequence encoding a membrane domain). The modified polynucleotides canbe chemically modified and/or structurally modified. When thepolynucleotides of the present disclosure are chemically and/orstructurally modified the polynucleotides can be referred to as“modified polynucleotides.”

The present disclosure provides for modified nucleosides and nucleotidesof a polynucleotide (e.g., RNA polynucleotides, such as mRNApolynucleotides) encoding a tethered IL-12 polypeptide. A “nucleoside”refers to a compound containing a sugar molecule (e.g., a pentose orribose) or a derivative thereof in combination with an organic base(e.g., a purine or pyrimidine) or a derivative thereof (also referred toherein as “nucleobase”). A “nucleotide” refers to a nucleoside includinga phosphate group. Modified nucleotides may by synthesized by any usefulmethod, such as, for example, chemically, enzymatically, orrecombinantly, to include one or more modified or non-naturalnucleosides. Polynucleotides may comprise a region or regions of linkednucleosides. Such regions may have variable backbone linkages. Thelinkages may be standard phosphodiester linkages, in which case thepolynucleotides would comprise regions of nucleotides.

The modified polynucleotides disclosed herein can comprise variousdistinct modifications. In some embodiments, the modifiedpolynucleotides contain one, two, or more (optionally different)nucleoside or nucleotide modifications. In some embodiments, a modifiedpolynucleotide, introduced to a cell may exhibit one or more desirableproperties, e.g., improved protein expression, reduced immunogenicity,or reduced degradation in the cell, as compared to an unmodifiedpolynucleotide.

a. Structural Modifications

In some embodiments, a polynucleotide of the present disclosure (e.g., apolynucleotide comprising a nucleotide sequence encoding an IL-12polypeptide and a nucleotide sequence encoding a membrane domain) isstructurally modified. As used herein, a “structural” modification isone in which two or more linked nucleosides are inserted, deleted,duplicated, inverted or randomized in a polynucleotide withoutsignificant chemical modification to the nucleotides themselves. Becausechemical bonds will necessarily be broken and reformed to effect astructural modification, structural modifications are of a chemicalnature and hence are chemical modifications. However, structuralmodifications will result in a different sequence of nucleotides. Forexample, the polynucleotide “ATCG” can be chemically modified to“AT-5meC-G”. The same polynucleotide can be structurally modified from“ATCG” to “ATCCCG (SEQ ID NO: 215)”. Here, the dinucleotide “CC” hasbeen inserted, resulting in a structural modification to thepolynucleotide.

b. Chemical Modifications

In some embodiments, the polynucleotides of the present disclosure(e.g., a polynucleotide comprising a nucleotide sequence encoding anIL-12 polypeptide and a nucleotide sequence encoding a membrane domain)are chemically modified. As used herein in reference to apolynucleotide, the terms “chemical modification” or, as appropriate,“chemically modified” refer to modification with respect to adenosine(A), guanosine (G), uridine (U), thymidine (T) or cytidine (C) ribo- ordeoxyribonucleosides in one or more of their position, pattern, percentor population, including, but not limited to, its nucleobase, sugar,backbone, or any combination thereof. Generally, herein, these terms arenot intended to refer to the ribonucleotide modifications in naturallyoccurring 5′-terminal mRNA cap moieties.

In some embodiments, the polynucleotides of the disclosure (e.g., apolynucleotide comprising a nucleotide sequence encoding an IL-12polypeptide and a nucleotide sequence encoding a membrane domain) canhave a uniform chemical modification of all or any of the samenucleoside type or a population of modifications produced by downwardtitration of the same starting modification in all or any of the samenucleoside type, or a measured percent of a chemical modification of allany of the same nucleoside type but with random incorporation, such aswhere all uridines are replaced by a uridine analog, e.g.,5-methoxyuridine. In another embodiment, the polynucleotides can have auniform chemical modification of two, three, or four of the samenucleoside type throughout the entire polynucleotide (such as alluridines and/or all cytidines, etc. are modified in the same way).

Modified nucleotide base pairing encompasses not only the standardadenosine-thymine, adenosine-uracil, or guanosine-cytosine base pairs,but also base pairs formed between nucleotides and/or modifiednucleotides comprising non-standard or modified bases, wherein thearrangement of hydrogen bond donors and hydrogen bond acceptors permitshydrogen bonding between a non-standard base and a standard base orbetween two complementary non-standard base structures. One example ofsuch non-standard base pairing is the base pairing between the modifiednucleotide inosine and adenine, cytosine or uracil. Any combination ofbase/sugar or linker may be incorporated into polynucleotides of thepresent disclosure.

The skilled artisan will appreciate that, except where otherwise noted,polynucleotide sequences set forth in the instant application willrecite “T”s in a representative DNA sequence but where the sequencerepresents RNA, the “T”s would be substituted for “U”s.

Modifications of polynucleotides (e.g., RNA polynucleotides, such asmRNA polynucleotides) that are useful in the composition of the presentdisclosure include, but are not limited to the following:2-methylthio-N6-(cis-hydroxyisopentenyl)adenosine;2-methylthio-N6-methyladenosine; 2-methylthio-N6-threonylcarbamoyladenosine; N6-glycinylcarbamoyladenosine;N6-isopentenyladenosine; N6-methyladenosine;N6-threonylcarbamoyladenosine; 1,2′-O-dimethyladenosine;1-methyladenosine; 2′-O-methyladenosine; 2′-O-ribosyladenosine(phosphate); 2-methyladenosine; 2-methylthio-N6 isopentenyladenosine;2-methylthio-N6-hydroxynorvalyl carbamoyladenosine;2′-O-methyladenosine; 2′-O-ribosyladenosine (phosphate);Isopentenyladenosine; N6-(cis-hydroxyisopentenyl)adenosine;N6,2′-O-dimethyladenosine; N6,2′-O-dimethyladenosine;N6,N6,2′-O-trimethyladenosine; N6,N6-dimethyladenosine;N6-acetyladenosine; N6-hydroxynorvalylcarbamoyladenosine;N6-methyl-N6-threonylcarbamoyladenosine; 2-methyladenosine;2-methylthio-N6-isopentenyladenosine; 7-deaza-adenosine;N1-methyl-adenosine; N6, N6 (dimethyl)adenine;N6-cis-hydroxy-isopentenyl-adenosine; α-thio-adenosine; 2(amino)adenine; 2 (aminopropyl)adenine; 2 (methylthio) N6(isopentenyl)adenine; 2-(alkyl)adenine; 2-(aminoalkyl)adenine;2-(aminopropyl)adenine; 2-(halo)adenine; 2-(halo)adenine;2-(propyl)adenine; 2′-Amino-2′-deoxy-ATP; 2′-Azido-2′-deoxy-ATP;2′-Deoxy-2′-α-aminoadenosine TP; 2′-Deoxy-2′-a-azidoadenosine TP; 6(alkyl)adenine; 6 (methyl)adenine; 6-(alkyl)adenine; 6-(methyl)adenine;7 (deaza)adenine; 8 (alkenyl)adenine; 8 (alkynyl)adenine; 8(amino)adenine; 8 (thioalkyl)adenine; 8-(alkenyl)adenine;8-(alkyl)adenine; 8-(alkynyl)adenine; 8-(amino)adenine; 8-(halo)adenine;8-(hydroxyl)adenine; 8-(thioalkyl)adenine; 8-(thiol)adenine;8-azido-adenosine; aza adenine; deaza adenine; N6 (methyl)adenine;N6-(isopentyl)adenine; 7-deaza-8-aza-adenosine; 7-methyladenine;1-Deazaadenosine TP; 2′Fluoro-N6-Bz-deoxyadenosine TP;2′-OMe-2-Amino-ATP; 2′O-methyl-N6-Bz-deoxyadenosine TP;2′-a-Ethynyladenosine TP; 2-aminoadenine; 2-Aminoadenosine TP;2-Amino-ATP; 2′-a-Trifluoromethyladenosine TP; 2-Azidoadenosine TP;2′-b-Ethynyladenosine TP; 2-Bromoadenosine TP;2′-b-Trifluoromethyladenosine TP; 2-Chloroadenosine TP;2′-Deoxy-2′,2′-difluoroadenosine TP; 2′-Deoxy-2′-a-mercaptoadenosine TP;2′-Deoxy-2′-a-thiomethoxyadenosine TP; 2′-Deoxy-2′-b-aminoadenosine TP;2′-Deoxy-2′-b-azidoadenosine TP; 2′-Deoxy-2′-b-bromoadenosine TP;2′-Deoxy-2′-b-chloroadenosine TP; 2′-Deoxy-2′-b-fluoroadenosine TP;2′-Deoxy-2′-b-iodoadenosine TP; 2′-Deoxy-2′-b-mercaptoadenosine TP;2′-Deoxy-2′-b-thiomethoxyadenosine TP; 2-Fluoroadenosine TP;2-Iodoadenosine TP; 2-Mercaptoadenosine TP; 2-methoxy-adenine;2-methylthio-adenine; 2-Trifluoromethyladenosine TP;3-Deaza-3-bromoadenosine TP; 3-Deaza-3-chloroadenosine TP;3-Deaza-3-fluoroadenosine TP; 3-Deaza-3-iodoadenosine TP;3-Deazaadenosine TP; 4′-Azidoadenosine TP; 4′-Carbocyclic adenosine TP;4′-Ethynyladenosine TP; 5′-Homo-adenosine TP; 8-Aza-ATP;8-bromo-adenosine TP; 8-Trifluoromethyladenosine TP; 9-DeazaadenosineTP; 2-aminopurine; 7-deaza-2,6-diaminopurine;7-deaza-8-aza-2,6-diaminopurine; 7-deaza-8-aza-2-aminopurine;2,6-diaminopurine; 7-deaza-8-aza-adenine, 7-deaza-2-aminopurine;2-thiocytidine; 3-methylcytidine; 5-formylcytidine;5-hydroxymethylcytidine; 5-methylcytidine; N4-acetylcytidine;2′-O-methylcytidine; 2′-O-methylcytidine; 5,2′-O-dimethylcytidine;5-formyl-2′-O-methylcytidine; Lysidine; N4,2′-O-dimethylcytidine;N4-acetyl-2′-O-methylcytidine; N4-methylcytidine;N4,N4-Dimethyl-2′-OMe-Cytidine TP; 4-methylcytidine; 5-aza-cytidine;Pseudo-iso-cytidine; pyrrolo-cytidine; α-thio-cytidine;2-(thio)cytosine; 2′-Amino-2′-deoxy-CTP; 2′-Azido-2′-deoxy-CTP;2′-Deoxy-2′-a-aminocytidine TP; 2′-Deoxy-2′-a-azidocytidine TP; 3(deaza) 5 (aza)cytosine; 3 (methyl)cytosine; 3-(alkyl)cytosine;3-(deaza) 5 (aza)cytosine; 3-(methyl)cytidine; 4,2′-O-dimethylcytidine;5 (halo)cytosine; 5 (methyl)cytosine; 5 (propynyl)cytosine; 5(trifluoromethyl)cytosine; 5-(alkyl)cytosine; 5-(alkynyl)cytosine;5-(halo)cytosine; 5-(propynyl)cytosine; 5-(trifluoromethyl)cytosine;5-bromo-cytidine; 5-iodo-cytidine; 5-propynyl cytosine; 6-(azo)cytosine;6-aza-cytidine; aza cytosine; deaza cytosine; N4 (acetyl)cytosine;1-methyl-1-deaza-pseudoisocytidine; 1-methyl-pseudoisocytidine;2-methoxy-5-methyl-cytidine; 2-methoxy-cytidine;2-thio-5-methyl-cytidine; 4-methoxy-1-methyl-pseudoisocytidine;4-methoxy-pseudoisocytidine; 4-thio-1-methyl-1-deaza-pseudoisocytidine;4-thio-1-methyl-pseudoisocytidine; 4-thio-pseudoisocytidine;5-aza-zebularine; 5-methyl-zebularine; pyrrolo-pseudoisocytidine;Zebularine; (E)-5-(2-Bromo-vinyl)cytidine TP; 2,2′-anhydro-cytidine TPhydrochloride; 2′Fluor-N4-Bz-cytidine TP; 2′Fluoro-N4-Acetyl-cytidineTP; 2′-O-Methyl-N4-Acetyl-cytidine TP; 2′O-methyl-N4-Bz-cytidine TP;2′-a-Ethynylcytidine TP; 2′-a-Trifluoromethylcytidine TP;2′-b-Ethynylcytidine TP; 2′-b-Trifluoromethylcytidine TP;2′-Deoxy-2′,2′-difluorocytidine TP; 2′-Deoxy-2′-a-mercaptocytidine TP;2′-Deoxy-2′-a-thiomethoxycytidine TP; 2′-Deoxy-2′-b-aminocytidine TP;2′-Deoxy-2′-b-azidocytidine TP; 2′-Deoxy-2′-b-bromocytidine TP;2′-Deoxy-2′-b-chlorocytidine TP; 2′-Deoxy-2′-b-fluorocytidine TP;2′-Deoxy-2′-b-iodocytidine TP; 2′-Deoxy-2′-b-mercaptocytidine TP;2′-Deoxy-2′-b-thiomethoxycytidine TP; 2′-O-Methyl-5-(1-propynyl)cytidineTP; 3′-Ethynylcytidine TP; 4′-Azidocytidine TP; 4′-Carbocyclic cytidineTP; 4′-Ethynylcytidine TP; 5-(1-Propynyl)ara-cytidine TP;5-(2-Chloro-phenyl)-2-thiocytidine TP; 5-(4-Amino-phenyl)-2-thiocytidineTP; 5-Aminoallyl-CTP; 5-Cyanocytidine TP; 5-Ethynylara-cytidine TP;5-Ethynylcytidine TP; 5′-Homo-cytidine TP; 5-Methoxycytidine TP;5-Trifluoromethyl-Cytidine TP; N4-Amino-cytidine TP; N4-Benzoyl-cytidineTP; Pseudoisocytidine; 7-methylguanosine; N2,2′-O-dimethylguanosine;N2-methylguanosine; Wyosine; 1,2′-O-dimethylguanosine;1-methylguanosine; 2′-O-methylguanosine; 2′-O-ribosylguanosine(phosphate); 2′-O-methylguanosine; 2′-O-ribosylguanosine (phosphate);7-aminomethyl-7-deazaguanosine; 7-cyano-7-deazaguanosine; Archaeosine;Methylwyosine; N2,7-dimethylguanosine; N2,N2,2′-O-trimethylguanosine;N2,N2,7-trimethylguanosine; N2,N2-dimethylguanosine;N2,7,2′-O-trimethylguanosine; 6-thio-guanosine; 7-deaza-guanosine;8-oxo-guanosine; N1-methyl-guanosine; α-thio-guanosine; 2(propyl)guanine; 2-(alkyl)guanine; 2′-Amino-2′-deoxy-GTP;2′-Azido-2′-deoxy-GTP; 2′-Deoxy-2′-a-aminoguanosine TP;2′-Deoxy-2′-a-azidoguanosine TP; 6 (methyl)guanine; 6-(alkyl)guanine;6-(methyl)guanine; 6-methyl-guanosine; 7 (alkyl)guanine; 7(deaza)guanine; 7 (methyl)guanine; 7-(alkyl)guanine; 7-(deaza)guanine;7-(methyl)guanine; 8 (alkyl)guanine; 8 (alkynyl)guanine; 8(halo)guanine; 8 (thioalkyl)guanine; 8-(alkenyl)guanine;8-(alkyl)guanine; 8-(alkynyl)guanine; 8-(amino)guanine; 8-(halo)guanine;8-(hydroxyl)guanine; 8-(thioalkyl)guanine; 8-(thiol)guanine; azaguanine; deaza guanine; N (methyl)guanine; N-(methyl)guanine;1-methyl-6-thio-guanosine; 6-methoxy-guanosine;6-thio-7-deaza-8-aza-guanosine; 6-thio-7-deaza-guanosine;6-thio-7-methyl-guanosine; 7-deaza-8-aza-guanosine;7-methyl-8-oxo-guanosine; N2,N2-dimethyl-6-thio-guanosine;N2-methyl-6-thio-guanosine; 1-Me-GTP; 2′Fluoro-N2-isobutyl-guanosine TP;2′O-methyl-N2-isobutyl-guanosine TP; 2′-a-Ethynylguanosine TP;2′-a-Trifluoromethylguanosine TP; 2′-b-Ethynylguanosine TP;2′-b-Trifluoromethylguanosine TP; 2′-Deoxy-2′,2′-difluoroguanosine TP;2′-Deoxy-2′-a-mercaptoguanosine TP; 2′-Deoxy-2′-a-thiomethoxyguanosineTP; 2′-Deoxy-2′-b-aminoguanosine TP; 2′-Deoxy-2′-b-azidoguanosine TP;2′-Deoxy-2′-b-bromoguanosine TP; 2′-Deoxy-2′-b-chloroguanosine TP;2′-Deoxy-2′-b-fluoroguanosine TP; 2′-Deoxy-2′-b-iodoguanosine TP;2′-Deoxy-2′-b-mercaptoguanosine TP; 2′-Deoxy-2′-b-thiomethoxyguanosineTP; 4′-Azidoguanosine TP; 4′-Carbocyclic guanosine TP;4′-Ethynylguanosine TP; 5′-Homo-guanosine TP; 8-bromo-guanosine TP;9-Deazaguanosine TP; N2-isobutyl-guanosine TP; 1-methylinosine; Inosine;1,2′-O-dimethylinosine; 2′-O-methylinosine; 7-methylinosine;2′-O-methylinosine; Epoxyqueuosine; galactosyl-queuosine;Mannosylqueuosine; Queuosine; allyamino-thymidine; aza thymidine; deazathymidine; deoxy-thymidine; 2′-O-methyluridine; 2-thiouridine;3-methyluridine; 5-carboxymethyluridine; 5-hydroxyuridine;5-methyluridine; 5-taurinomethyl-2-thiouridine; 5-taurinomethyluridine;Dihydrouridine; Pseudouridine; (3-(3-amino-3-carboxypropyl)uridine;1-methyl-3-(3-amino-5-carboxypropyl)pseudouridine;1-methylpseduouridine; 1-methyl-pseudouridine; 2′-O-methyluridine;2′-O-methylpseudouridine; 2′-O-methyluridine; 2-thio-2′-O-methyluridine;3-(3-amino-3-carboxypropyl)uridine; 3,2′-O-dimethyluridine;3-Methyl-pseudo-Uridine TP; 4-thiouridine;5-(carboxyhydroxymethyl)uridine; 5-(carboxyhydroxymethyl)uridine methylester; 5,2′-O-dimethyluridine; 5,6-dihydro-uridine;5-aminomethyl-2-thiouridine; 5-carbamoylmethyl-2′-O-methyluridine;5-carbamoylmethyluridine; 5-carboxyhydroxymethyluridine;5-carboxyhydroxymethyluridine methyl ester;5-carboxymethylaminomethyl-2′-O-methyluridine;5-carboxymethylaminomethyl-2-thiouridine;5-carboxymethylaminomethyl-2-thiouridine;5-carboxymethylaminomethyluridine; 5-carboxymethylaminomethyluridine;5-Carbamoylmethyluridine TP; 5-methoxycarbonylmethyl-2′-O-methyluridine;5-methoxycarbonylmethyl-2-thiouridine; 5-methoxycarbonylmethyluridine;5-methyluridine), 5-methoxyuridine; 5-methyl-2-thiouridine;5-methylaminomethyl-2-selenouridine; 5-methylaminomethyl-2-thiouridine;5-methylaminomethyluridine; 5-Methyldihydrouridine; 5-Oxyaceticacid-Uridine TP; 5-Oxyacetic acid-methyl ester-Uridine TP;N1-methyl-pseudo-uridine; uridine 5-oxyacetic acid; uridine 5-oxyaceticacid methyl ester; 3-(3-Amino-3-carboxypropyl)-Uridine TP;5-(iso-Pentenylaminomethyl)-2-thiouridine TP;5-(iso-Pentenylaminomethyl)-2′-O-methyluridine TP;5-(iso-Pentenylaminomethyl)uridine TP; 5-propynyl uracil;a-τhio-uridine; 1(aminoalkylamino-carbonylethylenyl)-2(thio)-pseudouracil; 1(aminoalkylaminocarbonylethylenyl)-2,4-(dithio)pseudouracil; 1(aminoalkylaminocarbonylethylenyl)-4 (thio)pseudouracil; 1(aminoalkylaminocarbonylethylenyl)-pseudouracil; 1(aminocarbonylethylenyl)-2(thio)-pseudouracil; 1(aminocarbonylethylenyl)-2,4-(dithio)pseudouracil; 1(aminocarbonylethylenyl)-4 (thio)pseudouracil; 1(aminocarbonylethylenyl)-pseudouracil; 1 substituted2(thio)-pseudouracil; 1 substituted 2,4-(dithio)pseudouracil; 1substituted 4 (thio)pseudouracil; 1 substituted pseudouracil;1-(aminoalkylamino-carbonylethylenyl)-2-(thio)-pseudouracil;1-Methyl-3-(3-amino-3-carboxypropyl) pseudouridine TP;1-Methyl-3-(3-amino-3-carboxypropyl)pseudo-UTP; 1-Methyl-pseudo-UTP; 2(thio)pseudouracil; 2′ deoxy uridine; 2′ fluorouridine; 2-(thio)uracil;2,4-(dithio)psuedouracil; 2′ methyl, 2′amino, 2′azido,2′fluro-guanosine; 2′-Amino-2′-deoxy-UTP; 2′-Azido-2′-deoxy-UTP;2′-Azido-deoxyuridine TP; 2′-O-methylpseudouridine; 2′ deoxy uridine; 2′fluorouridine; 2′-Deoxy-2′-a-aminouridine TP; 2′-Deoxy-2′-a-azidouridineTP; 2-methylpseudouridine; 3 (3 amino-3 carboxypropyl)uracil; 4(thio)pseudouracil; 4-(thio)pseudouracil; 4-(thio)uracil; 4-thiouracil;5 (1,3-diazole-1-alkyl)uracil; 5 (2-aminopropyl)uracil; 5(aminoalkyl)uracil; 5 (dimethylaminoalkyl)uracil; 5(guanidiniumalkyl)uracil; 5 (methoxycarbonylmethyl)-2-(thio)uracil; 5(methoxycarbonyl-methyl)uracil; 5 (methyl) 2 (thio)uracil; 5 (methyl)2,4 (dithio)uracil; 5 (methyl) 4 (thio)uracil; 5 (methylaminomethyl)-2(thio)uracil; 5 (methylaminomethyl)-2,4 (dithio)uracil; 5 (methylaminomethyl)-4 (thio)uracil; 5 (propynyl)uracil; 5(trifluoromethyl)uracil; 5-(2-aminopropyl)uracil;5-(alkyl)-2-(thio)pseudouracil; 5-(alkyl)-2,4 (dithio)pseudouracil;5-(alkyl)-4 (thio)pseudouracil; 5-(alkyl)pseudouracil; 5-(alkyl)uracil;5-(alkynyl)uracil; 5-(allylamino)uracil; 5-(cyanoalkyl)uracil;5-(dialkylaminoalkyl)uracil; 5-(dimethylaminoalkyl)uracil;5-(guanidiniumalkyl)uracil; 5-(halo)uracil;5-(1,3-diazole-1-alkyl)uracil; 5-(methoxy)uracil;5-(methoxycarbonylmethyl)-2-(thio)uracil;5-(methoxycarbonyl-methyl)uracil; 5-(methyl) 2(thio)uracil; 5-(methyl)2,4 (dithio)uracil; 5-(methyl) 4 (thio)uracil;5-(methyl)-2-(thio)pseudouracil; 5-(methyl)-2,4 (dithio)pseudouracil;5-(methyl)-4 (thio)pseudouracil; 5-(methyl)pseudouracil;5-(methylaminomethyl)-2 (thio)uracil;5-(methylaminomethyl)-2,4(dithio)uracil;5-(methylaminomethyl)-4-(thio)uracil; 5-(propynyl)uracil;5-(trifluoromethyl)uracil; 5-aminoallyl-uridine; 5-bromo-uridine;5-iodo-uridine; 5-uracil; 6 (azo)uracil; 6-(azo)uracil; 6-aza-uridine;allyamino-uracil; aza uracil; deaza uracil; N3 (methyl)uracil;Pseudo-UTP-1-2-ethanoic acid; Pseudouracil; 4-Thio-pseudo-UTP;1-carboxymethyl-pseudouridine; 1-methyl-1-deaza-pseudouridine;1-propynyl-uridine; 1-taurinomethyl-1-methyl-uridine;1-taurinomethyl-4-thio-uridine; 1-taurinomethyl-pseudouridine;2-methoxy-4-thio-pseudouridine; 2-thio-1-methyl-1-deaza-pseudouridine;2-thio-1-methyl-pseudouridine; 2-thio-5-aza-uridine;2-thio-dihydropseudouridine; 2-thio-dihydrouridine;2-thio-pseudouridine; 4-methoxy-2-thio-pseudouridine;4-methoxy-pseudouridine; 4-thio-1-methyl-pseudouridine;4-thio-pseudouridine; 5-aza-uridine; Dihydropseudouridine;(±)1-(2-Hydroxypropyl)pseudouridine TP;(2R)-1-(2-Hydroxypropyl)pseudouridine TP;(2S)-1-(2-Hydroxypropyl)pseudouridine TP;(E)-5-(2-Bromo-vinyl)ara-uridine TP; (E)-5-(2-Bromo-vinyl)uridine TP;(Z)-5-(2-Bromo-vinyl)ara-uridine TP; (Z) (2-Bromo-vinyl)uridine TP;1-(2,2,2-Trifluoroethyl)-pseudo-UTP;1-(2,2,3,3,3-Pentafluoropropyl)pseudouridine TP;1-(2,2-Diethoxyethyl)pseudouridine TP;1-(2,4,6-Trimethylbenzyl)pseudouridine TP;1-(2,4,6-Trimethyl-benzyl)pseudo-UTP;1-(2,4,6-Trimethyl-phenyl)pseudo-UTP;1-(2-Amino-2-carboxyethyl)pseudo-UTP; 1-(2-Amino-ethyl)pseudo-UTP;1-(2-Hydroxyethyl)pseudouridine TP; 1-(2-Methoxyethyl)pseudouridine TP;1-(3,4-Bis-trifluoromethoxybenzyl)pseudouridine TP;1-(3,4-Dimethoxybenzyl)pseudouridine TP;1-(3-Amino-3-carboxypropyl)pseudo-UTP; 1-(3-Amino-propyl)pseudo-UTP;1-(3-Cyclopropyl-prop-2-ynyl)pseudouridine TP;1-(4-Amino-4-carboxybutyl)pseudo-UTP; 1-(4-Amino-benzyl)pseudo-UTP;1-(4-Amino-butyl)pseudo-UTP; 1-(4-Amino-phenyl)pseudo-UTP;1-(4-Azidobenzyl)pseudouridine TP; 1-(4-Bromobenzyl)pseudouridine TP;1-(4-Chlorobenzyl)pseudouridine TP; 1-(4-Fluorobenzyl)pseudouridine TP;1-(4-Iodobenzyl)pseudouridine TP;1-(4-Methanesulfonylbenzyl)pseudouridine TP;1-(4-Methoxybenzyl)pseudouridine TP; 1-(4-Methoxy-benzyl)pseudo-UTP;1-(4-Methoxy-phenyl)pseudo-UTP; 1-(4-Methylbenzyl)pseudouridine TP;1-(4-Methyl-benzyl)pseudo-UTP; 1-(4-Nitrobenzyl)pseudouridine TP;1-(4-Nitro-benzyl)pseudo-UTP; 1(4-Nitro-phenyl)pseudo-UTP;1-(4-Thiomethoxybenzyl)pseudouridine TP;1-(4-Trifluoromethoxybenzyl)pseudouridine TP;1-(4-Trifluoromethylbenzyl)pseudouridine TP;1-(5-Amino-pentyl)pseudo-UTP; 1-(6-Amino-hexyl)pseudo-UTP;1,6-Dimethyl-pseudo-UTP;1-[3-(2-{2-[2-(2-Aminoethoxy)-ethoxy]-ethoxy}-ethoxy)-propionyl]pseudouridineTP; 1-[3-(2-(2-Aminoethoxy)-ethoxy]-propionyl pseudouridine TP;1-Acetylpseudouridine TP; 1-Alkyl-6-(1-propynyl)-pseudo-UTP;1-Alkyl-6-(2-propynyl)-pseudo-UTP; 1-Alkyl-6-allyl-pseudo-UTP;1-Alkyl-6-ethynyl-pseudo-UTP; 1-Alkyl-6-homoallyl-pseudo-UTP;1-Alkyl-6-vinyl-pseudo-UTP; 1-Allylpseudouridine TP;1-Aminomethyl-pseudo-UTP; 1-Benzoylpseudouridine TP;1-Benzyloxymethylpseudouridine TP; 1-Benzyl-pseudo-UTP;1-Biotinyl-PEG2-pseudouridine TP; 1-Biotinylpseudouridine TP;1-Butyl-pseudo-UTP; 1-Cyanomethylpseudouridine TP;1-Cyclobutylmethyl-pseudo-UTP; 1-Cyclobutyl-pseudo-UTP;1-Cycloheptylmethyl-pseudo-UTP; 1-Cycloheptyl-pseudo-UTP;1-Cyclohexylmethyl-pseudo-UTP; 1-Cyclohexyl-pseudo-UTP;1-Cyclooctylmethyl-pseudo-UTP; 1-Cyclooctyl-pseudo-UTP;1-Cyclopentylmethyl-pseudo-UTP; 1-Cyclopentyl-pseudo-UTP;1-Cyclopropylmethyl-pseudo-UTP; 1-Cyclopropyl-pseudo-UTP;1-Ethyl-pseudo-UTP; 1-Hexyl-pseudo-UTP; 1-Homoallylpseudouridine TP;1-Hydroxymethylpseudouridine TP; 1-iso-propyl-pseudo-UTP;1-Me-2-thio-pseudo-UTP; 1-Me-4-thio-pseudo-UTP;1-Me-alpha-thio-pseudo-UTP; 1-Methanesulfonylmethylpseudouridine TP;1-Methoxymethylpseudouridine TP;1-Methyl-6-(2,2,2-Trifluoroethyl)pseudo-UTP;1-Methyl-6-(4-morpholino)-pseudo-UTP;1-Methyl-6-(4-thiomorpholino)-pseudo-UTP; 1-Methyl-6-(substitutedphenyl)pseudo-UTP; 1-Methyl-6-amino-pseudo-UTP;1-Methyl-6-azido-pseudo-UTP; 1-Methyl-6-bromo-pseudo-UTP;1-Methyl-6-butyl-pseudo-UTP; 1-Methyl-6-chloro-pseudo-UTP;1-Methyl-6-cyano-pseudo-UTP; 1-Methyl-6-dimethylamino-pseudo-UTP;1-Methyl-6-ethoxy-pseudo-UTP; 1-Methyl-6-ethylcarboxylate-pseudo-UTP;1-Methyl-6-ethyl-pseudo-UTP; 1-Methyl-6-fluoro-pseudo-UTP;1-Methyl-6-formyl-pseudo-UTP; 1-Methyl-6-hydroxyamino-pseudo-UTP;1-Methyl-6-hydroxy-pseudo-UTP; 1-Methyl-6-iodo-pseudo-UTP;1-Methyl-6-iso-propyl-pseudo-UTP; 1-Methyl-6-methoxy-pseudo-UTP;1-Methyl-6-methylamino-pseudo-UTP; 1-Methyl-6-phenyl-pseudo-UTP;1-Methyl-6-propyl-pseudo-UTP; 1-Methyl-6-tert-butyl-pseudo-UTP;1-Methyl-6-trifluoromethoxy-pseudo-UTP;1-Methyl-6-trifluoromethyl-pseudo-UTP; 1-MorpholinomethylpseudouridineTP; 1-Pentyl-pseudo-UTP; 1-Phenyl-pseudo-UTP; 1-PivaloylpseudouridineTP; 1-Propargylpseudouridine TP; 1-Propyl-pseudo-UTP;1-propynyl-pseudouridine; 1-p-tolyl-pseudo-UTP; 1-tert-Butyl-pseudo-UTP;1-Thiomethoxymethylpseudouridine TP; 1-ThiomorpholinomethylpseudouridineTP; 1-Trifluoroacetylpseudouridine TP; 1-Trifluoromethyl-pseudo-UTP;1-Vinylpseudouridine TP; 2,2′-anhydro-uridine TP; 2′-bromo-deoxyuridineTP; 2′-F-5-Methyl-2′-deoxy-UTP; 2′-OMe-S-Me-UTP; 2′-OMe-pseudo-UTP;2′-a-Ethynyluridine TP; 2′-a-Trifluoromethyluridine TP;2′-b-Ethynyluridine TP; 2′-b-Trifluoromethyluridine TP;2′-Deoxy-2′,2′-difluorouridine TP; 2′-Deoxy-2′-a-mercaptouridine TP;2′-Deoxy-2′-a-thiomethoxyuridine TP; 2′-Deoxy-2′-b-aminouridine TP;2′-Deoxy-2′-b-azidouridine TP; 2′-Deoxy-2′-b-bromouridine TP;2′-Deoxy-2′-b-chlorouridine TP; 2′-Deoxy-2′-b-fluorouridine TP;2′-Deoxy-2′-b-iodouridine TP; 2′-Deoxy-2′-b-mercaptouridine TP;2′-Deoxy-2′-b-thiomethoxyuridine TP; 2-methoxy-4-thio-uridine;2-methoxyuridine; 2′-O-Methyl-5-(1-propynyl)uridine TP;3-Alkyl-pseudo-UTP; 4′-Azidouridine TP; 4′-Carbocyclic uridine TP;4′-Ethynyluridine TP; 5-(1-Propynyl)ara-uridine TP; 5-(2-Furanyl)uridineTP; 5-Cyanouridine TP; 5-Dimethylaminouridine TP; 5′-Homo-uridine TP;5-iodo-2′-fluoro-deoxyuridine TP; 5-Phenylethynyluridine TP;5-Trideuteromethyl-6-deuterouridine TP; 5-Trifluoromethyl-Uridine TP;5-Vinylarauridine TP; 6-(2,2,2-Trifluoroethyl)-pseudo-UTP;6-(4-Morpholino)-pseudo-UTP; 6-(4-Thiomorpholino)-pseudo-UTP;6-(Substituted-Phenyl)-pseudo-UTP; 6-Amino-pseudo-UTP;6-Azido-pseudo-UTP; 6-Bromo-pseudo-UTP; 6-Butyl-pseudo-UTP;6-Chloro-pseudo-UTP; 6-Cyano-pseudo-UTP; 6-Dimethylamino-pseudo-UTP;6-Ethoxy-pseudo-UTP; 6-Ethylcarboxylate-pseudo-UTP; 6-Ethyl-pseudo-UTP;6-Fluoro-pseudo-UTP; 6-Formyl-pseudo-UTP; 6-Hydroxyamino-pseudo-UTP;6-Hydroxy-pseudo-UTP; 6-Iodo-pseudo-UTP; 6-iso-Propyl-pseudo-UTP;6-Methoxy-pseudo-UTP; 6-Methylamino-pseudo-UTP; 6-Methyl-pseudo-UTP;6-Phenyl-pseudo-UTP; 6-Phenyl-pseudo-UTP; 6-Propyl-pseudo-UTP;6-tert-Butyl-pseudo-UTP; 6-Trifluoromethoxy-pseudo-UTP;6-Trifluoromethyl-pseudo-UTP; Alpha-thio-pseudo-UTP; Pseudouridine1-(4-methylbenzenesulfonic acid) TP; Pseudouridine 1-(4-methylbenzoicacid) TP; Pseudouridine TP 1-[3-(2-ethoxy)] propionic acid;Pseudouridine TP1-[3-{2-(2-[2-(2-ethoxy)-ethoxy]-ethoxy)-ethoxy}]propionic acid;Pseudouridine TP1-[3-{2-(2-[2-{2(2-ethoxy)-ethoxy}-ethoxy]-ethoxy)-ethoxy}]propionicacid; Pseudouridine TP 1-[3-{2-(2-[2-ethoxy]-ethoxy)-ethoxy}]propionicacid; Pseudouridine TP 1-[3-{2-(2-ethoxy)-ethoxy}] propionic acid;Pseudouridine TP 1-methylphosphonic acid; Pseudouridine TP1-methylphosphonic acid diethyl ester; Pseudo-UTP-N1-3-propionic acid;Pseudo-UTP-N1-4-butanoic acid; Pseudo-UTP-N1-5-pentanoic acid;Pseudo-UTP-N1-6-hexanoic acid; Pseudo-UTP-N1-7-heptanoic acid;Pseudo-UTP-N1-methyl-p-benzoic acid; Pseudo-UTP-N1-p-benzoic acid;Wybutosine; Hydroxywybutosine; Isowyosine; Peroxywybutosine;undermodified hydroxywybutosine; 4-demethylwyosine; 2, 6-(diamino)purine; 1-(aza)-2-(thio)-3-(aza)-phenoxazin-1-yl:1,3-(diaza)-2-(oxo)-phenthiazin-1-yl;1,3-(diaza)-2-(oxo)-phenoxazin-1-yl;1,3,5-(triaza)-2,6-(dioxa)-naphthalene; 2 (amino)purine;2,4,5-(trimethyl)phenyl; 2′ methyl, 2′amino, 2′azido, 2′fluro-cytidine;2′ methyl, 2′amino, 2′azido, 2′fluro-adenine; 2′methyl, 2′amino,2′azido, 2′fluro-uridine; 2′-amino-2′-deoxyribose;2-amino-6-Chloro-purine; 2-aza-inosinyl; 2′-azido-2′-deoxyribose;2′fluoro-2′-deoxyribose; 2′-fluoro-modified bases; 2′-O-methyl-ribose;2-oxo-7-aminopyridopyrimidin-3-yl; 2-oxo-pyridopyrimidine-3-yl;2-pyridinone; 3 nitropyrrole; 3-(methyl)-7-(propynyl)isocarbostyrilyl;3-(methyl)isocarbostyrilyl; 4-(fluoro)-6-(methyl)benzimidazole;4-(methyl)benzimidazole; 4-(methyl)indolyl; 4,6-(dimethyl)indolyl; 5nitroindole; 5 substituted pyrimidines; 5-(methyl)isocarbostyrilyl;5-nitroindole; 6-(aza)pyrimidine; 6-(azo)thymine;6-(methyl)-7-(aza)indolyl; 6-chloro-purine;6-phenyl-pyrrolo-pyrimidin-2-on-3-yl;7-(aminoalkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenthiazin-1-yl;7-(aminoalkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenoxazin-1-yl;7-(aminoalkylhydroxy)-1,3-(diaza)-2-(oxo)-phenoxazin-1-yl;7-(aminoalkylhydroxy)-1,3-(diaza)-2-(oxo)-phenthiazin-1-yl;7-(aminoalkylhydroxy)-1,3-(diaza)-2-(oxo)-phenoxazin-1-yl;7-(aza)indolyl;7-(guanidiniumalkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenoxazinl-yl;7-(guanidiniumalkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenthiazin-1-yl;7-(guanidiniumalkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenoxazin-1-yl;7-(guanidiniumalkylhydroxy)-1,3-(diaza)-2-(oxo)-phenoxazin-1-yl;7-(guanidiniumalkyl-hydroxy)-1,3-(diaza)-2-(oxo)-phenthiazin-1-yl;7-(guanidiniumalkylhydroxy)-1,3-(diaza)-2-(oxo)-phenoxazin-1-yl;7-(propynyl)isocarbostyrilyl; 7-(propynyl)isocarbostyrilyl,propynyl-7-(aza)indolyl; 7-deaza-inosinyl; 7-substituted1-(aza)-2-(thio)-3-(aza)-phenoxazin-1-yl; 7-substituted1,3-(diaza)-2-(oxo)-phenoxazin-1-yl; 9-(methyl)-imidizopyridinyl;Aminoindolyl; Anthracenyl;bis-ortho-(aminoalkylhydroxy)-6-phenyl-pyrrolo-pyrimidin-2-on-3-yl; bis-ortho-substituted-6-phenyl-pyrrolo-pyrimidin-2-on-3-yl; Difluorotolyl;Hypoxanthine; Imidizopyridinyl; Inosinyl; Isocarbostyrilyl;Isoguanisine; N2-substituted purines; N6-methyl-2-amino-purine;N6-substituted purines; N-alkylated derivative; Napthalenyl;Nitrobenzimidazolyl; Nitroimidazolyl; Nitroindazolyl; Nitropyrazolyl;Nubularine; 06-substituted purines; O-alkylated derivative;ortho-(aminoalkylhydroxy)-6-phenyl-pyrrolo-pyrimidin-2-on-3-yl;ortho-substituted-6-phenyl-pyrrolo-pyrimidin-2-on-3-yl; Oxoformycin TP;para-(aminoalkylhydroxy)-6-phenyl-pyrrolo-pyrimidin-2-on-3-yl;para-substituted-6-phenyl-pyrrolo-pyrimidin-2-on-3-yl; Pentacenyl;Phenanthracenyl; Phenyl; propynyl-7-(aza)indolyl; Pyrenyl;pyridopyrimidin-3-yl; pyridopyrimidin-3-yl,2-oxo-7-amino-pyridopyrimidin-3-yl; pyrrolo-pyrimidin-2-on-3-yl;Pyrrolopyrimidinyl; Pyrrolopyrizinyl; Stilbenzyl; substituted1,2,4-triazoles; Tetracenyl; Tubercidine; Xanthine; Xanthosine-5′-TP;2-thio-zebularine; 5-aza-2-thio-zebularine; 7-deaza-2-amino-purine;pyridin-4-one ribonucleoside; 2-Amino-riboside-TP; Formycin A TP;Formycin B TP; Pyrrolosine TP; 2′-OH-ara-adenosine TP;2′-OH-ara-cytidine TP; 2′-OH-ara-uridine TP; 2′-OH-ara-guanosine TP;5-(2-carbomethoxyvinyl)uridine TP; andN6-(19-Amino-pentaoxanonadecyl)adenosine TP.

In some embodiments, the polynucleotide (e.g., RNA polynucleotide, suchas mRNA polynucleotide) includes a combination of at least two (e.g., 2,3, 4 or more) of the aforementioned modified nucleobases.

In some embodiments, the mRNA comprises at least one chemically modifiednucleoside. In some embodiments, the at least one chemically modifiednucleoside is selected from the group consisting of pseudouridine (w),N1-methylpseudouridine (m1ψ), 2-thiouridine (s2U), 4′-thiouridine,5-methyl cytosine, 2-thio-1-methyl-1-deaza-pseudouridine,2-thio-1-methyl-pseudouridine, 2-thio-5-aza-uridine,2-thio-dihydropseudouridine, 2-thio-dihydrouridine,2-thio-pseudouridine, 4-methoxy-2-thio-pseudouridine,4-methoxy-pseudouridine, 4-thio-1-methyl-pseudouridine,4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine,5-methyluridine, 5-methoxyuridine, 2′-O-methyl uridine,1-methyl-pseudouridine (mlw), 5-methoxy-uridine (mo5U),5-methyl-cytidine (m5C), α-thio-guanosine, a-thio-adenosine, 5-cyanouridine, 4′-thio uridine 7-deaza-adenine, 1-methyl-adenosine (m1A),2-methyl-adenine (m2A), N6-methyl-adenosine (m6A), and2,6-Diaminopurine, (I), 1-methyl-inosine (mil), wyosine (imG),methylwyosine (mimG), 7-deaza-guanosine, 7-cyano-7-deaza-guanosine(preQ0), 7-aminomethyl-7-deaza-guanosine (preQ1), 7-methyl-guanosine(m7G), 1-methyl-guanosine (m1 G), 8-oxo-guanosine,7-methyl-8-oxo-guanosine, 2,8-dimethyladenosine, 2-geranylthiouridine,2-lysidine, 2-selenouridine,3-(3-amino-3-carboxypropyl)-5,6-dihydrouridine,3-(3-amino-3-carboxypropyl)pseudouridine, 3-methylpseudouridine,5-(carboxyhydroxymethyl)-2′-O-methyluridine methyl ester,5-aminomethyl-2-geranylthiouridine, 5-aminomethyl-2-selenouridine,5-aminomethyluridine, 5-carbamoylhydroxymethyluridine,5-carbamoylmethyl-2-thiouridine, 5-carboxymethyl-2-thiouridine,5-carboxymethylaminomethyl-2-geranylthiouridine,5-carboxymethylaminomethyl-2-selenouridine, 5-cyanomethyluridine,5-hydroxycytidine, 5-methylaminomethyl-2-geranylthiouridine,7-aminocarboxypropyl-demethylwyosine, 7-aminocarboxypropylwyosine,7-aminocarboxypropylwyosine methyl ester, 8-methyladenosine,N4,N4-dimethylcytidine, N6-formyladenosine, N6-hydroxymethyladenosine,agmatidine, cyclic N6-threonylcarbamoyladenosine, glutamyl-queuosine,methylated undermodified hydroxywybutosine,N4,N4,2′-O-trimethylcytidine, geranylated5-methylaminomethyl-2-thiouridine, geranylated5-carboxymethylaminomethyl-2-thiouridine, Qbase, preQ0base, preQ1base,and two or more combinations thereof. In some embodiments, the at leastone chemically modified nucleoside is selected from the group consistingof pseudouridine, N1-methylpseudouridine, 5-methylcytosine,5-methoxyuridine, and a combination thereof. In some embodiments, thepolynucleotide (e.g., RNA polynucleotide, such as mRNA polynucleotide)includes a combination of at least two (e.g., 2, 3, 4 or more) of theaforementioned modified nucleobases.

(i) Base Modifications

In certain embodiments, the chemical modification is at nucleobases inthe polynucleotides (e.g., RNA polynucleotide, such as mRNApolynucleotide).

In some embodiments, a polynucleotide as disclosed herein comprises atleast one chemically modified nucleobase.

In some embodiments, the at least one chemically modified nucleobase isselected from the group consisting of pseudouracil (w),N1-methylpseudouracil (m1ψ), 2-thiouracil (s2U), 4′-thiouracil,5-methylcytosine, 2-thio-1-methyl-1-deaza-pseudouracil,2-thio-1-methyl-pseudouracil, 2-thio-5-aza-uracil,2-thio-dihydropseudouracil, 2-thio-dihydrouracil, 2-thio-pseudouracil,4-methoxy-2-thio-pseudouracil, 4-methoxy-pseudouracil,4-thio-1-methyl-pseudouracil, 4-thio-pseudouracil, 5-aza-uracil,dihydropseudouracil, 5-methyluracil, 5-methoxyuracil, 2′-O-methyluracil, 1-methyl-pseudouracil (m1ψ), 5-methoxy-uracil (mo5U),5-methyl-cytosine (m5C), α-thio-guanine, α-thio-adenine, 5-cyano uracil,4′-thio uracil, 7-deaza-adenine, 1-methyl-adenine (m1A),2-methyl-adenine (m2A), N6-methyl-adenine (m6A), and 2,6-Diaminopurine,(I), 1-methyl-inosine (m1I), wyosine (imG), methylwyosine (mimG),7-deaza-guanine, 7-cyano-7-deaza-guanine (preQ0),7-aminomethyl-7-deaza-guanine (preQ1), 7-methyl-guanine (m7G),1-methyl-guanine (m1 G), 8-oxo-guanine, 7-methyl-8-oxo-guanine, and twoor more combinations thereof.

In some embodiments, the nucleobases in a polynucleotide as disclosedherein are chemically modified by at least about 10%, at least about20%, at least about 30%, at least about 40%, at least about 50%, atleast about 60%, at least about 70%, at least about 80%, at least about90%, at least 95%, at least 99%, or 100%.

In some embodiments, the chemically modified nucleobases are selectedfrom the group consisting of uracil, adenine, cytosine, guanine, and anycombination thereof.

In some embodiments, the uracils in a polynucleotide disclosed hereinare chemically modified by at least about 10%, at least about 20%, atleast about 30%, at least about 40%, at least about 50%, at least about60%, at least about 70%, at least about 80%, at least about 90%, atleast 95%, at least 99%, or 100%.

In some embodiments, the adenines in a polynucleotide disclosed hereinare chemically modified by at least about 10%, at least about 20%, atleast about 30%, at least about 40%, at least about 50%, at least about60%, at least about 70%, at least about 80%, at least about 90%, atleast 95%, at least 99%, or 100%.

In some embodiments, the cytosines in a polynucleotide disclosed hereinare chemically modified by at least about 10%, at least about 20%, atleast about 30%, at least about 40%, at least about 50%, at least about60%, at least about 70%, at least about 80%, at least about 90%, atleast 95%, at least 99%, or 100%.

In some embodiments, the guanines in a polynucleotide disclosed hereinare chemically modified by at least about 10%, at least about 20%, atleast about 30%, at least about 40%, at least about 50%, at least about60%, at least about 70%, at least about 80%, at least about 90%, atleast 95%, at least 99%, or 100%.

In some embodiments, the polynucleotide (e.g., RNA polynucleotide, suchas mRNA polynucleotide) includes a combination of at least two (e.g., 2,3, 4 or more) of modified nucleobases. In some embodiments, modifiednucleobases in the polynucleotide (e.g., RNA polynucleotide, such asmRNA polynucleotide) are selected from the group consisting of1-methyl-pseudouridine (m1ψ), 5-methoxy-uridine (mo5U),5-methyl-cytidine (m5C), pseudouridine (ψ), α-thio-guanosine andα-thio-adenosine. In some embodiments, the polynucleotide includes acombination of at least two (e.g., 2, 3, 4 or more) of theaforementioned modified nucleobases.

In some embodiments, the polynucleotide (e.g., RNA polynucleotide, suchas mRNA polynucleotide) comprises pseudouridine (w) and5-methyl-cytidine (m5C). In some embodiments, the polynucleotide (e.g.,RNA polynucleotide, such as mRNA polynucleotide) comprises1-methyl-pseudouridine (m1ψ). In some embodiments, the polynucleotide(e.g., RNA polynucleotide, such as mRNA polynucleotide) comprises1-methyl-pseudouridine (m1ψ) and 5-methyl-cytidine (m5C). In someembodiments, the polynucleotide (e.g., RNA polynucleotide, such as mRNApolynucleotide) comprises 2-thiouridine (s2U). In some embodiments, thepolynucleotide (e.g., RNA polynucleotide, such as mRNA polynucleotide)comprises 2-thiouridine and 5-methyl-cytidine (m5C). In someembodiments, the polynucleotide (e.g., RNA polynucleotide, such as mRNApolynucleotide) comprises methoxy-uridine (mo5U). In some embodiments,the polynucleotide (e.g., RNA polynucleotide, such as mRNApolynucleotide) comprises 5-methoxy-uridine (mo5U) and 5-methyl-cytidine(m5C). In some embodiments, the polynucleotide (e.g., RNApolynucleotide, such as mRNA polynucleotide) comprises 2′-O-methyluridine. In some embodiments, the polynucleotide (e.g., RNApolynucleotide, such as mRNA polynucleotide) comprises 2′-O-methyluridine and 5-methyl-cytidine (m5C). In some embodiments, thepolynucleotide (e.g., RNA polynucleotide, such as mRNA polynucleotide)comprises N6-methyl-adenosine (m6A). In some embodiments, thepolynucleotide (e.g., RNA polynucleotide, such as mRNA polynucleotide)comprises N6-methyl-adenosine (m6A) and 5-methyl-cytidine (m5C).

In some embodiments, the polynucleotide (e.g., RNA polynucleotide, suchas mRNA polynucleotide) is uniformly modified (e.g., fully modified,modified throughout the entire sequence) for a particular modification.For example, a polynucleotide can be uniformly modified with5-methyl-cytidine (m5C), meaning that all cytosine residues in the mRNAsequence are replaced with 5-methyl-cytidine (m5C). Similarly, apolynucleotide can be uniformly modified for any type of nucleosideresidue present in the sequence by replacement with a modified residuesuch as any of those set forth above.

In some embodiments, the modified nucleobase is a modified cytosine.Examples of nucleobases and nucleosides having a modified cytosineinclude N4-acetyl-cytidine (ac4C), 5-methyl-cytidine (m5C),5-halo-cytidine (e.g., 5-iodo-cytidine), 5-hydroxymethyl-cytidine(hm5C), 1-methyl-pseudoisocytidine, 2-thio-cytidine (s2C), and2-thio-5-methyl-cytidine.

In some embodiments, a modified nucleobase is a modified uridine.Example nucleobases and nucleosides having a modified uridine include5-cyano uridine or 4′-thio uridine.

In some embodiments, a modified nucleobase is a modified adenine.Example nucleobases and nucleosides having a modified adenine include7-deaza-adenine, 1-methyl-adenosine (m1A), 2-methyl-adenine (m2A),N6-methyl-adenine (m6A), and 2,6-Diaminopurine.

In some embodiments, a modified nucleobase is a modified guanine.Example nucleobases and nucleosides having a modified guanine includeinosine (I), 1-methyl-inosine (mil), wyosine (imG), methylwyosine(mimG), 7-deaza-guanosine, 7-cyano-7-deaza-guanosine (preQ0),7-aminomethyl-7-deaza-guanosine (preQ1), 7-methyl-guanosine (m7G),1-methyl-guanosine (m1 G), 8-oxo-guanosine, 7-methyl-8-oxo-guanosine.

In some embodiments, the nucleobases, sugar, backbone, or anycombination thereof in a polynucleotide, nucleic acid sequence, and/orORF as disclosed herein are chemically modified by at least 10%, atleast 20%, at least 30%, at least 40%, at least 50%, at least 60%, atleast 70%, at least 80%, at least 90%, at least 95%, at least 99%, or100%.

In some embodiments, the polynucleotides can include any useful linkerbetween the nucleosides, including subunit linkers, membrane domainlinkers, and heterologous polypeptide linkers as disclosed elsewhereherein. Such linkers, including backbone modifications, that are usefulin the composition of the present disclosure include, but are notlimited to the following: 3′-alkylene phosphonates, 3′-aminophosphoramidate, alkene containing backbones,aminoalkylphosphoramidates, aminoalkylphosphotriesters,boranophosphates, —CH₂—O—N(CH₃)—CH₂—, —CH₂—N(CH₃)—N(CH₃)—CH₂—,—CH₂—NH—CH₂—, chiral phosphonates, chiral phosphorothioates, formacetyland thioformacetyl backbones, methylene (methylimino), methyleneformacetyl and thioformacetyl backbones, methyleneimino andmethylenehydrazino backbones, morpholino linkages, —N(CH₃)—CH₂—CH₂—,oligonucleosides with heteroatom internucleoside linkage, phosphinates,phosphoramidates, phosphorodithioates, phosphorothioate internucleosidelinkages, phosphorothioates, phosphotriesters, PNA, siloxane backbones,sulfamate backbones, sulfide sulfoxide and sulfone backbones, sulfonateand sulfonamide backbones, thionoalkylphosphonates,thionoalkylphosphotriesters, and thionophosphoramidates.

(ii) Sugar Modifications

The modified nucleosides and nucleotides (e.g., building blockmolecules), which can be incorporated into a polynucleotide (e.g., RNAor mRNA, as described herein), can be modified on the sugar of theribonucleic acid. For example, the 2′ hydroxyl group (OH) can bemodified or replaced with a number of different substituents. Exemplarysubstitutions at the 2′-position include, but are not limited to, H,halo, optionally substituted Ci-6 alkyl; optionally substituted Ci-6alkoxy; optionally substituted C₆₋₁₀ aryloxy; optionally substitutedC₃-8 cycloalkyl; optionally substituted C₃-8 cycloalkoxy; optionallysubstituted C₆₋₁₀ aryloxy; optionally substituted C₆₋₁₀ aryl-C₁₋₆alkoxy, optionally substituted Ci-12 (heterocyclyl)oxy; a sugar (e.g.,ribose, pentose, or any described herein); a polyethyleneglycol (PEG),—O(CH₂CH₂O)_(n)CH₂CH₂OR, where R is H or optionally substituted alkyl,and n is an integer from 0 to 20 (e.g., from 0 to 4, from 0 to 8, from 0to 10, from 0 to 16, from 1 to 4, from 1 to 8, from 1 to 10, from 1 to16, from 1 to 20, from 2 to 4, from 2 to 8, from 2 to 10, from 2 to 16,from 2 to 20, from 4 to 8, from 4 to 10, from 4 to 16, and from 4 to20); “locked” nucleic acids (LNA) in which the 2′-hydroxyl is connectedby a Ci-6 alkylene or Ci-6 heteroalkylene bridge to the 4′-carbon of thesame ribose sugar, where exemplary bridges included methylene,propylene, ether, or amino bridges; aminoalkyl, as defined herein;aminoalkoxy, as defined herein; amino as defined herein; and amino acid,as defined herein

Generally, RNA includes the sugar group ribose, which is a 5-memberedring having an oxygen. Exemplary, non-limiting modified nucleotidesinclude replacement of the oxygen in ribose (e.g., with S, Se, oralkylene, such as methylene or ethylene); addition of a double bond(e.g., to replace ribose with cyclopentenyl or cyclohexenyl); ringcontraction of ribose (e.g., to form a 4-membered ring of cyclobutane oroxetane); ring expansion of ribose (e.g., to form a 6- or 7-memberedring having an additional carbon or heteroatom, such as foranhydrohexitol, altritol, mannitol, cyclohexanyl, cyclohexenyl, andmorpholino that also has a phosphoramidate backbone); multicyclic forms(e.g., tricyclo; and “unlocked” forms, such as glycol nucleic acid (GNA)(e.g., R-GNA or S-GNA, where ribose is replaced by glycol units attachedto phosphodiester bonds), threose nucleic acid (TNA, where ribose isreplace with α-L-threofuranosyl-(3′→2)), and peptide nucleic acid (PNA,where 2-amino-ethyl-glycine linkages replace the ribose andphosphodiester backbone). The sugar group can also contain one or morecarbons that possess the opposite stereochemical configuration than thatof the corresponding carbon in ribose. Thus, a polynucleotide moleculecan include nucleotides containing, e.g., arabinose, as the sugar. Suchsugar modifications are taught International Patent Publication Nos.WO2013052523 and WO2014093924, the contents of each of which areincorporated herein by reference in their entireties.

(iii) Combinations of Modifications

The polynucleotides of the disclosure (e.g., a polynucleotide comprisinga nucleotide sequence encoding a tethered IL-12 polypeptide) can includea combination of modifications to the sugar, the nucleobase, and/or theinternucleoside linkage. These combinations can include any one or moremodifications described herein.

Combinations of modified nucleotides can be used to form thepolynucleotides of the disclosure. Unless otherwise noted, the modifiednucleotides can be completely substituted for the natural nucleotides ofthe polynucleotides of the disclosure. As a non-limiting example, thenatural nucleotide uridine can be substituted with a modified nucleosidedescribed herein. In another non-limiting example, the naturalnucleotide uridine can be partially substituted or replaced (e.g., about0.1%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%,65%, 70%, 75%, 80%, 85%, 90%, 95% or 99.9%) with at least one of themodified nucleoside disclosed herein. Any combination of base/sugar orlinker can be incorporated into the polynucleotides of the disclosureand such modifications are taught in International Patent PublicationsWO2013052523 and WO2014093924, and U.S. Publ. Nos. US 20130115272 andUS20150307542, the contents of each of which are incorporated herein byreference in its entirety.

13. Untranslated Regions (UTRs)

Untranslated regions (UTRs) are nucleic acid sections of apolynucleotide before a start codon (5′UTR) and after a stop codon(3′UTR) that are not translated. In some embodiments, a polynucleotide(e.g., a ribonucleic acid (RNA), e.g., a messenger RNA (mRNA)) of thedisclosure further comprises UTR (e.g., a 5′UTR or functional fragmentthereof, a 3′UTR or functional fragment thereof, or a combinationthereof).

A UTR can be homologous or heterologous to the coding region in apolynucleotide. In some embodiments, the UTR is homologous to thenucleic acid sequence encoding the IL-12 polypeptide. In someembodiments, the UTR is heterologous to the nucleic acide sequenceencoding the IL-12 polypeptide. In some embodiments, the polynucleotidecomprises two or more 5′UTRs or functional fragments thereof, each ofwhich have the same or different nucleotide sequences. In someembodiments, the polynucleotide comprises two or more 3′UTRs orfunctional fragments thereof, each of which have the same or differentnucleotide sequences.

In some embodiments, the 5′UTR or functional fragment thereof, 3′ UTR orfunctional fragment thereof, or any combination thereof is sequenceoptimized.

In some embodiments, the 5′UTR or functional fragment thereof, 3′ UTR orfunctional fragment thereof, or any combination thereof comprises atleast one chemically modified nucleobase, e.g., 1 methylpseudouridine or5-methoxyuracil.

UTRs can have features that provide a regulatory role, e.g., increasedor decreased stability, localization and/or translation efficiency. Apolynucleotide comprising a UTR can be administered to a cell, tissue,or organism, and one or more regulatory features can be measured usingroutine methods. In some embodiments, a functional fragment of a 5′UTRor 3′UTR comprises one or more regulatory features of a full length 5′or 3′ UTR, respectively.

Natural 5′UTRs bear features that play roles in translation initiation.They harbor signatures like Kozak sequences that are commonly known tobe involved in the process by which the ribosome initiates translationof many genes. Kozak sequences have the consensus CCR(A/G)CCAUGG (SEQ IDNO: 216), where R is a purine (adenine or guanine) three bases upstreamof the start codon (AUG), which is followed by another ‘G’. 5′UTRs alsohave been known to form secondary structures that are involved inelongation factor binding.

By engineering the features typically found in abundantly expressedgenes of specific target organs, one can enhance the stability andprotein production of a polynucleotide. For example, introduction of5′UTR of liver-expressed mRNA, such as albumin, serum amyloid A,Apolipoprotein AB/E, transferrin, alpha fetoprotein, erythropoietin, orFactor VIII, can enhance expression of polynucleotides in hepatic celllines or liver. Likewise, use of 5′UTR from other tissue-specific mRNAto improve expression in that tissue is possible for muscle (e.g., MyoD,Myosin, Myoglobin, Myogenin, Herculin), for endothelial cells (e.g.,Tie-1, CD36), for myeloid cells (e.g., C/EBP, AML1, G-CSF, GM-CSF,CD11b, MSR, Fr-1, i-NOS), for leukocytes (e.g., CD45, CD18), for adiposetissue (e.g., CD36, GLUT4, ACRP30, adiponectin) and for lung epithelialcells (e.g., SP-A/B/C/D).

In some embodiments, UTRs are selected from a family of transcriptswhose proteins share a common function, structure, feature or property.For example, an encoded polypeptide can belong to a family of proteins(i.e., that share at least one function, structure, feature,localization, origin, or expression pattern), which are expressed in aparticular cell, tissue or at some time during development. The UTRsfrom any of the genes or mRNA can be swapped for any other UTR of thesame or different family of proteins to create a new polynucleotide.

In some embodiments, the 5′UTR and the 3′UTR can be heterologous. Insome embodiments, the 5′UTR can be derived from a different species thanthe 3′UTR. In some embodiments, the 3′UTR can be derived from adifferent species than the 5′UTR.

Co-owned International Patent Application No. PCT/US2014/021522 (Publ.No. WO/2014/164253, incorporated herein by reference in its entirety)provides a listing of exemplary UTRs that can be utilized in thepolynucleotide of the present disclosure as flanking regions to an ORF.

Exemplary UTRs of the application include, but are not limited to, oneor more 5′UTR and/or 3′UTR derived from the nucleic acid sequence of: aglobin, such as an α- or β-globin (e.g., a Xenopus, mouse, rabbit, orhuman globin); a strong Kozak translational initiation signal; a CYBA(e.g., human cytochrome b-245 α polypeptide); an albumin (e.g., humanalbumin7); a HSD17B4 (hydroxysteroid (17-β) dehydrogenase); a virus(e.g., a tobacco etch virus (TEV), a Venezuelan equine encephalitisvirus (VEEV), a Dengue virus, a cytomegalovirus (CMV) (e.g., CMVimmediate early 1 (IE1)), a hepatitis virus (e.g., hepatitis B virus), asindbis virus, or a PAV barley yellow dwarf virus); a heat shock protein(e.g., hsp70); a translation initiation factor (e.g., elF4G); a glucosetransporter (e.g., hGLUT1 (human glucose transporter 1)); an actin(e.g., human α or β actin); a GAPDH; a tubulin; a histone; a citric acidcycle enzyme; a topoisomerase (e.g., a 5′UTR of a TOP gene lacking the5′ TOP motif (the oligopyrimidine tract)); a ribosomal protein Large 32(L32); a ribosomal protein (e.g., human or mouse ribosomal protein, suchas, for example, rps9); an ATP synthase (e.g., ATP5A1 or the β subunitof mitochondrial H⁺-ATP synthase); a growth hormone e (e.g., bovine(bGH) or human (hGH)); an elongation factor (e.g., elongation factor 1al (EEF1A1)); a manganese superoxide dismutase (MnSOD); a myocyteenhancer factor 2A (MEF2A); a β-F1-ATPase, a creatine kinase, amyoglobin, a granulocyte-colony stimulating factor (G-CSF); a collagen(e.g., collagen type I, alpha 2 (Col1A2), collagen type I, alpha 1(Col1A1), collagen type VI, alpha 2 (Col6A2), collagen type VI, alpha 1(Col6A1)); a ribophorin (e.g., ribophorin I (RPNI)); a low densitylipoprotein receptor-related protein (e.g., LRP1); a cardiotrophin-likecytokine factor (e.g., Nnt1); calreticulin (Calr); a procollagen-lysine,2-oxoglutarate 5-dioxygenase 1 (Plodl); and a nucleobindin (e.g.,Nucbl).

In some embodiments, the 5′UTR is selected from the group consisting ofa β-globin 5′UTR; a 5′UTR containing a strong Kozak translationalinitiation signal; a cytochrome b-245 α polypeptide (CYBA) 5′UTR; ahydroxysteroid (1713) dehydrogenase (HSD17B4) 5′UTR; a Tobacco etchvirus (TEV) 5′UTR; a Venezuelen equine encephalitis virus (TEEV) 5′UTR;a 5′ proximal open reading frame of rubella virus (RV) RNA encodingnonstructural proteins; a Dengue virus (DEN) 5′UTR; a heat shock protein70 (Hsp70) 5′UTR; a eIF4G 5′UTR; a GLUT1 5′UTR; functional fragmentsthereof and any combination thereof.

In some embodiments, the 3′UTR is selected from the group consisting ofa β-globin 3′UTR; a CYBA 3′UTR; an albumin 3′UTR; a growth hormone (GH)3′UTR; a VEEV 3′UTR; a hepatitis B virus (HBV) 3′UTR; a-μlobin 3′UTR; aDEN 3′UTR; a PAV barley yellow dwarf virus (BYDV-PAV) 3′UTR; anelongation factor 1 al (EEF1A1) 3′UTR; a manganese superoxide dismutase(MnSOD) 3′UTR; a f3 subunit of mitochondrial H(+)-ATP synthase (β-mRNA)3′UTR; a GLUT1 3′UTR; a MEF2A 3′UTR; a β-F1-ATPase 3′UTR; functionalfragments thereof and combinations thereof.

Wild-type UTRs derived from any gene or mRNA can be incorporated intothe polynucleotides of the disclosure. In some embodiments, a UTR can bealtered relative to a wild type or native UTR to produce a variant UTR,e.g., by changing the orientation or location of the UTR relative to theORF; or by inclusion of additional nucleotides, deletion of nucleotides,swapping or transposition of nucleotides. In some embodiments, variantsof 5′ or 3′ UTRs can be utilized, for example, mutants of wild typeUTRs, or variants wherein one or more nucleotides are added to orremoved from a terminus of the UTR.

Additionally, one or more synthetic UTRs can be used in combination withone or more non-synthetic UTRs. See, e.g., Mandal and Rossi, Nat.Protoc. 2013 8(3):568-82, the contents of which are incorporated hereinby reference in their entirety, and sequences available atwww.addgene.org/Derrick_Rossi/, last accessed Apr. 16, 2016. UTRs orportions thereof can be placed in the same orientation as in thetranscript from which they were selected or can be altered inorientation or location. Hence, a 5′ and/or 3′ UTR can be inverted,shortened, lengthened, or combined with one or more other 5′ UTRs or 3′UTRs.

In some embodiments, the polynucleotide comprises multiple UTRs, e.g., adouble, a triple or a quadruple 5′UTR or 3′UTR. For example, a doubleUTR comprises two copies of the same UTR either in series orsubstantially in series. For example, a double beta-globin 3′UTR can beused (see US2010/0129877, the contents of which are incorporated hereinby reference in its entirety). In certain embodiments, thepolynucleotides of the disclosure comprise a 5′UTR and/or a 3′UTRselected from any of the UTRs disclosed herein.

In some embodiments, the 5′UTR comprises a sequence selected from thegroup consisting of: SEQ ID NOs: 55-63 and 82-97.

In some embodiments, the 3′UTR comprises a sequence selected from thegroup consisting of: SEQ ID NOs: 64-81.

In certain embodiments, the 5′UTR and/or 3′UTR sequence of thedisclosure comprises a nucleotide sequence at least about 60%, at leastabout 70%, at least about 80%, at least about 90%, at least about 95%,at least about 96%, at least about 97%, at least about 98%, at leastabout 99%, or about 100% identical to a sequence selected from the groupconsisting of 5′UTR sequences comprising any of the 5′UTR sequencesdisclosed herein and/or 3′UTR sequences comprises any of the 3′UTRsequences disclosed herein, and any combination thereof.

In certain embodiments, the 3′ UTR sequence comprises one or more miRNAbinding sites, e.g., miR-122 binding sites, or any other heterologousnucleotide sequences therein, without disrupting the function of the 3′UTR. Some examples of 3′ UTR sequences comprising a miRNA binding siteare set forth in SEQ ID NOs: 222-224. In some embodiments, the 3′ UTRsequence comprises a nucleotide sequence at least about 60%, at leastabout 70%, at least about 80%, at least about 90%, at least about 95%,at least about 96%, at least about 97%, at least about 98%, at leastabout 99%, or about 100% identical to a sequence selected from the groupconsisting of SEQ ID NOs: 222-224. In certain embodiments, the 3′ UTRsequence comprises a nucleotide sequence at least about 60%, at leastabout 70%, at least about 80%, at least about 90%, at least about 95%,at least about 96%, at least about 97%, at least about 98%, at leastabout 99%, or about 100% identical to SEQ ID NO: 222. In certainembodiments, the 3′ UTR sequence comprises a nucleotide sequence atleast about 60%, at least about 70%, at least about 80%, at least about90%, at least about 95%, at least about 96%, at least about 97%, atleast about 98%, at least about 99%, or about 100% identical to SEQ IDNO: 223. In certain embodiments, the 3′ UTR sequence comprises anucleotide sequence at least about 60%, at least about 70%, at leastabout 80%, at least about 90%, at least about 95%, at least about 96%,at least about 97%, at least about 98%, at least about 99%, or about100% identical to SEQ ID NO: 224.

The polynucleotides of the disclosure can comprise combinations offeatures. For example, the ORF can be flanked by a 5′UTR that comprisesa strong Kozak translational initiation signal and/or a 3′UTR comprisingan oligo(dT) sequence for templated addition of a poly-A tail. A 5′UTRcan comprise a first polynucleotide fragment and a second polynucleotidefragment from the same and/or different UTRs (see, e.g., US2010/0293625,herein incorporated by reference in its entirety).

Other non-UTR sequences can be used as regions or subregions within thepolynucleotides of the disclosure. For example, introns or portions ofintron sequences can be incorporated into the polynucleotides of thedisclosure. Incorporation of intronic sequences can increase proteinproduction as well as polynucleotide expression levels. In someembodiments, the polynucleotide of the disclosure comprises an internalribosome entry site (IRES) instead of or in addition to a UTR (see,e.g., Yakubov et al., Biochem. Biophys. Res. Commun. 2010394(1):189-193, the contents of which are incorporated herein byreference in their entirety). In some embodiments, the polynucleotidecomprises an IRES instead of a 5′UTR sequence. In some embodiments, thepolynucleotide comprises an ORF and a viral capsid sequence. In someembodiments, the polynucleotide comprises a synthetic 5′UTR incombination with a non-synthetic 3′UTR.

In some embodiments, the UTR can also include at least one translationenhancer polynucleotide, translation enhancer element, or translationalenhancer elements (collectively, “TEE,” which refers to nucleic acidsequences that increase the amount of polypeptide or protein producedfrom a polynucleotide. As a non-limiting example, the TEE can includethose described in US2009/0226470, incorporated herein by reference inits entirety, and others known in the art. As a non-limiting example,the TEE can be located between the transcription promoter and the startcodon. In some embodiments, the 5′UTR comprises a TEE.

In one aspect, a TEE is a conserved element in a UTR that can promotetranslational activity of a nucleic acid such as, but not limited to,cap-dependent or cap-independent translation. In one non-limitingexample, the TEE comprises the TEE sequence in the 5′-leader of the Gtxhomeodomain protein. See Chappell et al., PNAS 2004 101:9590-9594,incorporated herein by reference in its entirety.

In some embodiments, the polynucleotide of the invention comprises oneor multiple copies of a TEE. The TEE in a translational enhancerpolynucleotide can be organized in one or more sequence segments. Asequence segment can harbor one or more of the TEEs provided herein,with each TEE being present in one or more copies. When multiplesequence segments are present in a translational enhancerpolynucleotide, they can be homogenous or heterogeneous. Thus, themultiple sequence segments in a translational enhancer polynucleotidecan harbor identical or different types of the TEE provided herein,identical or different number of copies of each of the TEE, and/oridentical or different organization of the TEE within each sequencesegment. In one embodiment, the polynucleotide of the inventioncomprises a translational enhancer polynucleotide sequence. Non-limitingexamples of TEE sequences are described in U.S. Publication2014/0200261, the contents of which are incorporated herein by referencein their entirety.

14. Functional RNA Elements

In some embodiments, the disclosure provides polynucleotides comprisinga modification (e.g., an RNA element), wherein the modification providesa desired translational regulatory activity. In some embodiments, thedisclosure provides a polynucleotide comprising a 5′ untranslated region(UTR), an initiation codon, a full open reading frame encoding apolypeptide, a 3′ UTR, and at least one modification, wherein the atleast one modification provides a desired translational regulatoryactivity, for example, a modification that promotes and/or enhances thetranslational fidelity of mRNA translation. In some embodiments, thedesired translational regulatory activity is a cis-acting regulatoryactivity. In some embodiments, the desired translational regulatoryactivity is an increase in the residence time of the 43S pre-initiationcomplex (PIC) or ribosome at, or proximal to, the initiation codon. Insome embodiments, the desired translational regulatory activity is anincrease in the initiation of polypeptide synthesis at or from theinitiation codon. In some embodiments, the desired translationalregulatory activity is an increase in the amount of polypeptidetranslated from the full open reading frame. In some embodiments, thedesired translational regulatory activity is an increase in the fidelityof initiation codon decoding by the PIC or ribosome. In someembodiments, the desired translational regulatory activity is inhibitionor reduction of leaky scanning by the PIC or ribosome. In someembodiments, the desired translational regulatory activity is a decreasein the rate of decoding the initiation codon by the PIC or ribosome. Insome embodiments, the desired translational regulatory activity isinhibition or reduction in the initiation of polypeptide synthesis atany codon within the mRNA other than the initiation codon. In someembodiments, the desired translational regulatory activity is inhibitionor reduction of the amount of polypeptide translated from any openreading frame within the mRNA other than the full open reading frame. Insome embodiments, the desired translational regulatory activity isinhibition or reduction in the production of aberrant translationproducts. In some embodiments, the desired translational regulatoryactivity is a combination of one or more of the foregoing translationalregulatory activities.

Accordingly, the present disclosure provides a polynucleotide, e.g., anmRNA, comprising an RNA element that comprises a sequence and/or an RNAsecondary structure(s) that provides a desired translational regulatoryactivity as described herein. In some aspects, the mRNA comprises an RNAelement that comprises a sequence and/or an RNA secondary structure(s)that promotes and/or enhances the translational fidelity of mRNAtranslation. In some aspects, the mRNA comprises an RNA element thatcomprises a sequence and/or an RNA secondary structure(s) that providesa desired translational regulatory activity, such as inhibiting and/orreducing leaky scanning. In some aspects, the disclosure provides anmRNA that comprises an RNA element that comprises a sequence and/or anRNA secondary structure(s) that inhibits and/or reduces leaky scanningthereby promoting the translational fidelity of the mRNA.

In some embodiments, the RNA element comprises natural and/or modifiednucleotides. In some embodiments, the RNA element comprises of asequence of linked nucleotides, or derivatives or analogs thereof, thatprovides a desired translational regulatory activity as describedherein. In some embodiments, the RNA element comprises a sequence oflinked nucleotides, or derivatives or analogs thereof, that forms orfolds into a stable RNA secondary structure, wherein the RNA secondarystructure provides a desired translational regulatory activity asdescribed herein. RNA elements can be identified and/or characterizedbased on the primary sequence of the element (e.g., GC-rich element), byRNA secondary structure formed by the element (e.g. stem-loop), by thelocation of the element within the RNA molecule (e.g., located withinthe 5′ UTR of an mRNA), by the biological function and/or activity ofthe element (e.g., “translational enhancer element”), and anycombination thereof.

In some embodiments, the disclosure provides an mRNA having one or morestructural modifications that inhibits leaky scanning and/or promotesthe translational fidelity of mRNA translation, wherein at least one ofthe structural modifications is a GC-rich RNA element. In someembodiments, the disclosure provides an mRNA comprising at least onemodification, wherein at least one modification is a GC-rich RNA elementcomprising a sequence of linked nucleotides, or derivatives or analogsthereof, preceding a Kozak consensus sequence in a 5′ UTR of the mRNA.In one embodiment, the GC-rich RNA element is located about 30, about25, about 20, about 15, about 10, about 5, about 4, about 3, about 2, orabout 1 nucleotide(s) upstream of a Kozak consensus sequence in the 5′UTR of the mRNA. In another embodiment, the GC-rich RNA element islocated 15-30, 15-20, 15-25, 10-15, or 5-10 nucleotides upstream of aKozak consensus sequence. In another embodiment, the GC-rich RNA elementis located immediately adjacent to a Kozak consensus sequence in the 5′UTR of the mRNA.

In some embodiments, the disclosure provides a GC-rich RNA element whichcomprises a sequence of 3-30, 5-25, 10-20, 15-20, about 20, about 15,about 12, about 10, about 7, about 6 or about 3 nucleotides, derivativesor analogs thereof, linked in any order, wherein the sequencecomposition is 70-80% cytosine, 60-70% cytosine, 50%-60% cytosine,40-50% cytosine, 30-40% cytosine bases. In some embodiments, thedisclosure provides a GC-rich RNA element which comprises a sequence of3-30, 5-25, 10-20, 15-20, about 20, about 15, about 12, about 10, about7, about 6 or about 3 nucleotides, derivatives or analogs thereof,linked in any order, wherein the sequence composition is about 80%cytosine, about 70% cytosine, about 60% cytosine, about 50% cytosine,about 40% cytosine, or about 30% cytosine.

In some embodiments, the disclosure provides a GC-rich RNA element whichcomprises a sequence of 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9,8, 7, 6, 5, 4, or 3 nucleotides, or derivatives or analogs thereof,linked in any order, wherein the sequence composition is 70-80%cytosine, 60-70% cytosine, 50%-60% cytosine, 40-50% cytosine, or 30-40%cytosine. In some embodiments, the disclosure provides a GC-rich RNAelement which comprises a sequence of 20, 19, 18, 17, 16, 15, 14, 13,12, 11, 10, 9, 8, 7, 6, 5, 4, or 3 nucleotides, or derivatives oranalogs thereof, linked in any order, wherein the sequence compositionis about 80% cytosine, about 70% cytosine, about 60% cytosine, about 50%cytosine, about 40% cytosine, or about 30% cytosine.

In some embodiments, the disclosure provides an mRNA comprising at leastone modification, wherein at least one modification is a GC-rich RNAelement comprising a sequence of linked nucleotides, or derivatives oranalogs thereof, preceding a Kozak consensus sequence in a 5′ UTR of themRNA, wherein the GC-rich RNA element is located about 30, about 25,about 20, about 15, about 10, about 5, about 4, about 3, about 2, orabout 1 nucleotide(s) upstream of a Kozak consensus sequence in the 5′UTR of the mRNA, and wherein the GC-rich RNA element comprises asequence of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,or 20 nucleotides, or derivatives or analogs thereof, linked in anyorder, wherein the sequence composition is >50% cytosine. In someembodiments, the sequence composition is >55% cytosine, >60%cytosine, >65% cytosine, >70% cytosine, >75% cytosine, >80%cytosine, >85% cytosine, or >90% cytosine.

In some embodiments, the disclosure provides an mRNA comprising at leastone modification, wherein at least one modification is a GC-rich RNAelement comprising a sequence of linked nucleotides, or derivatives oranalogs thereof, preceding a Kozak consensus sequence in a 5′ UTR of themRNA, wherein the GC-rich RNA element is located about 30, about 25,about 20, about 15, about 10, about 5, about 4, about 3, about 2, orabout 1 nucleotide(s) upstream of a Kozak consensus sequence in the 5′UTR of the mRNA, and wherein the GC-rich RNA element comprises asequence of about 3-30, 5-25, 10-20, 15-20 or about 20, about 15, about12, about 10, about 6 or about 3 nucleotides, or derivatives oranalogues thereof, wherein the sequence comprises a repeating GC-motif,wherein the repeating GC-motif is [CCG]n, wherein n=1 to 10, n=2 to 8,n=3 to 6, or n=4 to 5. In some embodiments, the sequence comprises arepeating GC-motif [CCG]n, wherein n=1, 2, 3, 4 or 5. In someembodiments, the sequence comprises a repeating GC-motif [CCG]n, whereinn=1, 2, or 3. In some embodiments, the sequence comprises a repeatingGC-motif [CCG]n, wherein n=1. In some embodiments, the sequencecomprises a repeating GC-motif [CCG]n, wherein n=2. In some embodiments,the sequence comprises a repeating GC-motif [CCG]n, wherein n=3. In someembodiments, the sequence comprises a repeating GC-motif [CCG]n, whereinn=4 (SEQ ID NO: 256). In some embodiments, the sequence comprises arepeating GC-motif [CCG]n, wherein n=5 (SEQ ID NO: 257).

In some embodiments, the GC-rich RNA element is located about 30, about25, about 20, about 15, about 10, about 5, about 4, about 3, about 2, orabout 1 nucleotide(s) upstream of a Kozak consensus sequence in the 5′UTR of the mRNA. In another embodiment, the GC-rich RNA element islocated about 15-30, 15-20, 15-25, 10-15, or 5-10 nucleotides upstreamof a Kozak consensus sequence. In another embodiment, the GC-rich RNAelement is located immediately adjacent to a Kozak consensus sequence inthe 5′ UTR of the mRNA.

In some embodiments, the disclosure provides an mRNA comprising at leastone modification, wherein at least one modification is a GC-rich RNAelement comprising the sequence set forth in SEQ ID NO: 258, orderivatives or analogs thereof, preceding a Kozak consensus sequence inthe 5′ UTR of the mRNA. In some embodiments, the GC-rich elementcomprises the sequence as set forth in SEQ ID NO: 258 locatedimmediately adjacent to and upstream of the Kozak consensus sequence inthe 5′ UTR of the mRNA. In some embodiments, the GC-rich elementcomprises the sequence as set forth in SEQ ID NO: 258 located 1, 2, 3,4, 5, 6, 7, 8, 9 or 10 bases upstream of the Kozak consensus sequence inthe 5′ UTR of the mRNA. In other embodiments, the GC-rich elementcomprises the sequence as set forth in SEQ ID NO: 258 located 1-3, 3-5,5-7, 7-9, 9-12, or 12-15 bases upstream of the Kozak consensus sequencein the 5′ UTR of the mRNA.

In some embodiments, the disclosure provides an mRNA comprising at leastone modification, wherein at least one modification is a GC-rich RNAelement comprising the sequence as set forth SEQ ID NO: 259, orderivatives or analogs thereof, preceding a Kozak consensus sequence inthe 5′ UTR of the mRNA. In some embodiments, the GC-rich elementcomprises the sequence as set forth SEQ ID NO: 259 located immediatelyadjacent to and upstream of the Kozak consensus sequence in the 5′ UTRof the mRNA. In some embodiments, the GC-rich element comprises thesequence as set forth SEQ ID NO: 259 located 1, 2, 3, 4, 5, 6, 7, 8, 9or 10 bases upstream of the Kozak consensus sequence in the 5′ UTR ofthe mRNA. In other embodiments, the GC-rich element comprises thesequence as set forth SEQ ID NO: 259 located 1-3, 3-5, 5-7, 7-9, 9-12,or 12-15 bases upstream of the Kozak consensus sequence in the 5′ UTR ofthe mRNA.

In some embodiments, the disclosure provides an mRNA comprising at leastone modification, wherein at least one modification is a GC-rich RNAelement comprising the sequence as set forth in SEQ ID NO: 260, orderivatives or analogs thereof, preceding a Kozak consensus sequence inthe 5′ UTR of the mRNA. In some embodiments, the GC-rich elementcomprises the sequence as set forth in SEQ ID NO: 260 locatedimmediately adjacent to and upstream of the Kozak consensus sequence inthe 5′ UTR of the mRNA. In some embodiments, the GC-rich elementcomprises the sequence as set forth in SEQ ID NO: 260 located 1, 2, 3,4, 5, 6, 7, 8, 9 or 10 bases upstream of the Kozak consensus sequence inthe 5′ UTR of the mRNA. In other embodiments, the GC-rich elementcomprises the sequence as set forth in SEQ ID NO: 260 located 1-3, 3-5,5-7, 7-9, 9-12, or 12-15 bases upstream of the Kozak consensus sequencein the 5′ UTR of the mRNA.

In some embodiments, the disclosure provides an mRNA comprising at leastone modification, wherein at least one modification is a GC-rich RNAelement comprising the sequence set forth in SEQ ID NO: 258, orderivatives or analogs thereof, preceding a Kozak consensus sequence inthe 5′ UTR of the mRNA, wherein the 5′ UTR comprises the sequence setforth in SEQ ID NO: 261.

In some embodiments, the GC-rich element comprises the sequence setforth in SEQ ID NO: 258 located immediately adjacent to and upstream ofthe Kozak consensus sequence in a 5′ UTR sequence described herein. Insome embodiments, the GC-rich element comprises the sequence set forthin SEQ ID NO: 258 located 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 bases upstreamof the Kozak consensus sequence in the 5′ UTR of the mRNA, wherein the5′ UTR comprises the sequence shown in SEQ ID NO: 261.

In other embodiments, the GC-rich element comprises the sequence setforth in SEQ ID NO: 258 located 1-3, 3-5, 5-7, 7-9, 9-12, or 12-15 basesupstream of the Kozak consensus sequence in the 5′ UTR of the mRNA,wherein the 5′ UTR comprises the sequence set forth in SEQ ID NO: 261.

In some embodiments, the 5′ UTR comprises the sequence set forth in SEQID NO: 262. In some embodiments, the 5′ UTR comprises the sequence setforth in SEQ ID NO: 263. In some embodiments, the disclosure provides anmRNA comprising at least one modification, wherein at least onemodification is a GC-rich RNA element comprising a stable RNA secondarystructure comprising a sequence of nucleotides, or derivatives oranalogs thereof, linked in an order which forms a hairpin or astem-loop. In one embodiment, the stable RNA secondary structure isupstream of the Kozak consensus sequence. In another embodiment, thestable RNA secondary structure is located about 30, about 25, about 20,about 15, about 10, or about 5 nucleotides upstream of the Kozakconsensus sequence. In another embodiment, the stable RNA secondarystructure is located about 20, about 15, about 10 or about 5 nucleotidesupstream of the Kozak consensus sequence. In another embodiment, thestable RNA secondary structure is located about 5, about 4, about 3,about 2, about 1 nucleotides upstream of the Kozak consensus sequence.In another embodiment, the stable RNA secondary structure is locatedabout 15-30, about 15-20, about 15-25, about 10-15, or about 5-10nucleotides upstream of the Kozak consensus sequence. In anotherembodiment, the stable RNA secondary structure is located 12-15nucleotides upstream of the Kozak consensus sequence. In anotherembodiment, the stable RNA secondary structure has a deltaG of about −30kcal/mol, about −20 to −30 kcal/mol, about −20 kcal/mol, about −10 to−20 kcal/mol, about −10 kcal/mol, about −5 to ˜10 kcal/mol.

In another embodiment, the modification is operably linked to an openreading frame encoding a polypeptide and wherein the modification andthe open reading frame are heterologous.

In another embodiment, the sequence of the GC-rich RNA element iscomprised exclusively of guanine (G) and cytosine (C) nucleobases.

RNA elements that provide a desired translational regulatory activity asdescribed herein can be identified and characterized using knowntechniques, such as ribosome profiling. Ribosome profiling is atechnique that allows the determination of the positions of PICs and/orribosomes bound to mRNAs (see e.g., Ingolia et al., (2009) Science324(5924):218-23, incorporated herein by reference). The technique isbased on protecting a region or segment of mRNA, by the PIC and/orribosome, from nuclease digestion. Protection results in the generationof a 30-bp fragment of RNA termed a ‘footprint’. The sequence andfrequency of RNA footprints can be analyzed by methods known in the art(e.g., RNA-seq). The footprint is roughly centered on the A-site of theribosome. If the PIC or ribosome dwells at a particular position orlocation along an mRNA, footprints generated at these position would berelatively common. Studies have shown that more footprints are generatedat positions where the PIC and/or ribosome exhibits decreasedprocessivity and fewer footprints where the PIC and/or ribosome exhibitsincreased processivity (Gardin et al., (2014) eLife 3:e03735). In someembodiments, residence time or the time of occupancy of a the PIC orribosome at a discrete position or location along an polynucleotidecomprising any one or more of the RNA elements described herein isdetermined by ribosome profiling.

15. MicroRNA (miRNA) Binding Sites

Sensor sequences include, for example, microRNA (miRNA) binding sites,transcription factor binding sites, structured mRNA sequences and/ormotifs, artificial binding sites engineered to act as pseudo-receptorsfor endogenous nucleic acid binding molecules, and combinations thereof.Non-limiting examples of sensor sequences are described in U.S.Publication 2014/0200261, the contents of which are incorporated hereinby reference in their entirety.

In some embodiments, a polynucleotide (e.g., a ribonucleic acid (RNA),e.g., a messenger RNA (mRNA)) of the disclosure further comprises asensor sequence. In some embodiments, the sensor sequence is a miRNAbinding site.

A miRNA is a 19-25 nucleotide long noncoding RNA that binds to apolynucleotide and down-regulates gene expression either by reducingstability or by inhibiting translation of the polynucleotide. A miRNAsequence comprises a “seed” region, i.e., a sequence in the region ofpositions 2-8 of the mature miRNA. A miRNA seed can comprise positions2-8 or 2-7 of the mature miRNA. In some embodiments, a miRNA seed cancomprise 7 nucleotides (e.g., nucleotides 2-8 of the mature miRNA),wherein the seed-complementary site in the corresponding miRNA bindingsite is flanked by an adenosine (A) opposed to miRNA position 1. In someembodiments, a miRNA seed can comprise 6 nucleotides (e.g., nucleotides2-7 of the mature miRNA), wherein the seed-complementary site in thecorresponding miRNA binding site is flanked by an adenosine (A) opposedto miRNA position 1. See, for example, Grimson A, Farh K K, Johnston WK, Garrett-Engele P, Lim L P, Bartel D P; Mol Cell. 2007 Jul. 6;27(1):91-105. miRNA profiling of the target cells or tissues can beconducted to determine the presence or absence of miRNA in the cells ortissues. In some embodiments, a polynucleotide (e.g., a ribonucleic acid(RNA), e.g., a messenger RNA (mRNA)) of the disclosure comprises one ormore microRNA target sequences, microRNA sequences, or microRNA seeds.Such sequences can correspond to any known microRNA such as those taughtin US Publication US2005/0261218 and US Publication US2005/0059005, thecontents of each of which are incorporated herein by reference in theirentirety.

As used herein, the term “microRNA (miRNA or miR) binding site” refersto a sequence within a polynucleotide, e.g., within a DNA or within anRNA transcript, including in the 5′UTR and/or 3′UTR, that has sufficientcomplementarity to all or a region of a miRNA to interact with,associate with or bind to the miRNA. In some embodiments, apolynucleotide of the disclosure further comprises a miRNA binding site.In exemplary embodiments, a 5′UTR and/or 3′UTR of the polynucleotide(e.g., a ribonucleic acid (RNA), e.g., a messenger RNA (mRNA)) comprisesa miRNA binding site.

A miRNA binding site having sufficient complementarity to a miRNA refersto a degree of complementarity sufficient to facilitate miRNA-mediatedregulation of a polynucleotide, e.g., miRNA-mediated translationalrepression or degradation of the polynucleotide. In exemplary aspects ofthe disclosure, a miRNA binding site having sufficient complementarityto the miRNA refers to a degree of complementarity sufficient tofacilitate miRNA-mediated degradation of the polynucleotide, e.g.,miRNA-guided RNA-induced silencing complex (RISC)-mediated cleavage ofmRNA. The miRNA binding site can have complementarity to, for example, a19-25 nucleotide miRNA sequence, to a 19-23 nucleotide miRNA sequence,or to a 22 nucleotide miRNA sequence. A miRNA binding site can becomplementary to only a portion of a miRNA, e.g., to a portion less than1, 2, 3, or 4 nucleotides of the full length of a naturally-occurringmiRNA sequence. Full or complete complementarity (e.g., fullcomplementarity or complete complementarity over all or a significantportion of the length of a naturally-occurring miRNA) is preferred whenthe desired regulation is mRNA degradation.

In some embodiments, a miRNA binding site includes a sequence that hascomplementarity (e.g., partial or complete complementarity) with a miRNAseed sequence. In some embodiments, the miRNA binding site includes asequence that has complete complementarity with a miRNA seed sequence.In some embodiments, a miRNA binding site includes a sequence that hascomplementarity (e.g., partial or complete complementarity) with a miRNAsequence. In some embodiments, the miRNA binding site includes asequence that has complete complementarity with a miRNA sequence. Insome embodiments, a miRNA binding site has complete complementarity witha miRNA sequence but for 1, 2, or 3 nucleotide substitutions, terminaladditions, and/or truncations.

In some embodiments, the miRNA binding site is the same length as thecorresponding miRNA. In other embodiments, the miRNA binding site isone, two, three, four, five, six, seven, eight, nine, ten, eleven ortwelve nucleotide(s) shorter than the corresponding miRNA at the 5′terminus, the 3′ terminus, or both. In still other embodiments, themicroRNA binding site is two nucleotides shorter than the correspondingmicroRNA at the 5′ terminus, the 3′ terminus, or both. The miRNA bindingsites that are shorter than the corresponding miRNAs are still capableof degrading the mRNA incorporating one or more of the miRNA bindingsites or preventing the mRNA from translation.

In some embodiments, the miRNA binding site binds to the correspondingmature miRNA that is part of an active RISC containing Dicer. In anotherembodiment, binding of the miRNA binding site to the corresponding miRNAin RISC degrades the mRNA containing the miRNA binding site or preventsthe mRNA from being translated. In some embodiments, the miRNA bindingsite has sufficient complementarity to miRNA so that a RISC complexcomprising the miRNA cleaves the polynucleotide comprising the miRNAbinding site. In other embodiments, the miRNA binding site has imperfectcomplementarity so that a RISC complex comprising the miRNA inducesinstability in the polynucleotide comprising the miRNA binding site. Inanother embodiment, the miRNA binding site has imperfect complementarityso that a RISC complex comprising the miRNA represses transcription ofthe polynucleotide comprising the miRNA binding site.

In some embodiments, the miRNA binding site has one, two, three, four,five, six, seven, eight, nine, ten, eleven or twelve mismatch(es) fromthe corresponding miRNA.

In some embodiments, the miRNA binding site has at least about ten, atleast about eleven, at least about twelve, at least about thirteen, atleast about fourteen, at least about fifteen, at least about sixteen, atleast about seventeen, at least about eighteen, at least about nineteen,at least about twenty, or at least about twenty-one contiguousnucleotides complementary to at least about ten, at least about eleven,at least about twelve, at least about thirteen, at least about fourteen,at least about fifteen, at least about sixteen, at least aboutseventeen, at least about eighteen, at least about nineteen, at leastabout twenty, or at least about twenty-one, respectively, contiguousnucleotides of the corresponding miRNA.

By engineering one or more miRNA binding sites into a polynucleotide ofthe disclosure, the polynucleotide can be targeted for degradation orreduced translation, provided the miRNA in question is available. Thiscan reduce off-target effects upon delivery of the polynucleotide. Forexample, if a polynucleotide of the disclosure is not intended to bedelivered to a tissue or cell but ends up there, then a miRNA abundantin the tissue or cell can inhibit the expression of the gene of interestif one or multiple binding sites of the miRNA are engineered into the5′UTR and/or 3′UTR of the polynucleotide.

Conversely, miRNA binding sites can be removed from polynucleotidesequences in which they naturally occur in order to increase proteinexpression in specific tissues. For example, a binding site for aspecific miRNA can be removed from a polynucleotide to improve proteinexpression in tissues or cells containing the miRNA.

In one embodiment, a polynucleotide of the disclosure can include atleast one miRNA-binding site in the 5′UTR and/or 3′UTR in order todirect cytotoxic or cytoprotective mRNA therapeutics to specific cellssuch as, but not limited to, normal and/or cancerous cells. In anotherembodiment, a polynucleotide of the disclosure can include two, three,four, five, six, seven, eight, nine, ten, or more miRNA-binding sites inthe 5′-UTR and/or 3′-UTR in order to direct cytotoxic or cytoprotectivemRNA therapeutics to specific cells such as, but not limited to, normaland/or cancerous cells.

Regulation of expression in multiple tissues can be accomplished throughintroduction or removal of one or more miRNA binding sites. The decisionwhether to remove or insert a miRNA binding site can be made based onmiRNA expression patterns and/or their profilings in diseases.Identification of miRNAs, miRNA binding sites, and their expressionpatterns and role in biology have been reported (e.g., Bonauer et al.,Curr Drug Targets 2010 11:943-949; Anand and Cheresh Curr Opin Hematol2011 18:171-176; Contreras and Rao Leukemia 2012 26:404-413 (2011 Dec.20. doi: 10.1038/leu.2011.356); Bartel Cell 2009 136:215-233; Landgrafet al, Cell, 2007 129:1401-1414; Gentner and Naldini, Tissue Antigens.2012 80:393-403 and all references therein; each of which isincorporated herein by reference in its entirety).

miRNAs and miRNA binding sites can correspond to any known sequence,including non-limiting examples described in U.S. Publication Nos.2014/0200261, 2005/0261218, and 2005/0059005, each of which areincorporated herein by reference in their entirety.

Examples of tissues where miRNA are known to regulate mRNA, and therebyprotein expression, include, but are not limited to, liver (miR-122),muscle (miR-133, miR-206, miR-208), endothelial cells (miR-17-92,miR-126), myeloid cells (miR-142-3p, miR-142-5p, miR-16, miR-21,miR-223, miR-24, miR-27), adipose tissue (let-7, miR-30c), heart(miR-1d, miR-149), kidney (miR-192, miR-194, miR-204), and lungepithelial cells (let-7, miR-133, miR-126).

Some microRNAs, e.g., miR-122, are abundant in normal tissue but arepresent in much lower levels in cancer or tumor tissue. Thus,engineering microRNA target sequences (i.e., microRNA binding site) intothe polynucleotides of the disclosure can effectively target themolecule for degradation or reduced translation in normal tissue (wherethe microRNA is abundant) while providing high levels of translation inthe cancer or tumor tissue (where the microRNA is present in much lowerlevels). This provides a tumor-targeting approach for the methods andcompositions of the disclosure.

Further examples of the miRNA binding sites that can be useful for thepresent disclosure include immune cell specific miRNAs including, butnot limited to, hsa-let-7a-2-3p, hsa-let-7a-3p, hsa-7a-5p, hsa-let-7c,hsa-let-7e-3p, hsa-let-7e-5p, hsa-let-7g-3p, hsa-let-7g-5p,hsa-let-7i-3p, hsa-let-7i-5p, miR-10a-3p, miR-10a-5p, miR-1184,hsa-let-7f-1-3p, hsa-let-7f-2-5p, hsa-let-7f-5p, miR-125b-1-3p,miR-125b-2-3p, miR-125b-5p, miR-1279, miR-130a-3p, miR-130a-5p,miR-132-3p, miR-132-5p, miR-142-3p, miR-142-5p, miR-143-3p, miR-143-5p,miR-146a-3p, miR-146a-5p, miR-146b-3p, miR-146b-5p, miR-147a, miR-147b,miR-148a-5p, miR-148a-3p, miR-150-3p, miR-150-5p, miR-151b, miR-155-3p,miR-155-5p, miR-15a-3p, miR-15a-5p, miR-15b-5p, miR-15b-3p, miR-16-1-3p,miR-16-2-3p, miR-16-5p, miR-17-5p, miR-181a-3p, miR-181a-5p,miR-181a-2-3p, miR-182-3p, miR-182-5p, miR-197-3p, miR-197-5p,miR-21-5p, miR-21-3p, miR-214-3p, miR-214-5p, miR-223-3p, miR-223-5p,miR-221-3p, miR-221-5p, miR-23b-3p, miR-23b-5p, miR-24-1-5p,miR-24-2-5p, miR-24-3p, miR-26a-1-3p, miR-26a-2-3p, miR-26a-5p,miR-26b-3p, miR-26b-5p, miR-27a-3p, miR-27a-5p, miR-27b-3p, miR-27b-5p,miR-28-3p, miR-28-5p, miR-2909, miR-29a-3p, miR-29a-5p, miR-29b-1-5p,miR-29b-2-5p, miR-29c-3p, miR-29c-5p, miR-30e-3p, miR-30e-5p,miR-331-5p, miR-339-3p, miR-339-5p, miR-345-3p, miR-345-5p, miR-346,miR-34a-3p, miR-34a-5p, miR-363-3p, miR-363-5p, miR-372, miR-377-3p,miR-377-5p, miR-493-3p, miR-493-5p, miR-542, miR-548b-5p, miR548c-5p,miR-548i, miR-548j, miR-548n, miR-574-3p, miR-598, miR-718, miR-935,miR-99a-3p, miR-99a-5p, miR-99b-3p, and miR-99b-5p. Furthermore, novelmiRNAs can be identified in immune cell through micro-arrayhybridization and microtome analysis (e.g., Jima D D et al, Blood, 2010,116:e118-e127; Vaz C et al., BMC Genomics, 2010, 11,288, the content ofeach of which is incorporated herein by reference in its entirety.)

MiRNAs that are known to be expressed in the liver include, but are notlimited to, miR-107, miR-122-3p, miR-122-5p, miR-1228-3p, miR-1228-5p,miR-1249, miR-129-5p, miR-1303, miR-151a-3p, miR-151a-5p, miR-152,miR-194-3p, miR-194-5p, miR-199a-3p, miR-199a-5p, miR-199b-3p,miR-199b-5p, miR-296-5p, miR-557, miR-581, miR-939-3p, and miR-939-5p.MiRNA binding sites from any liver specific miRNA can be introduced toor removed from a polynucleotide of the disclosure to regulateexpression of the polynucleotide in the liver. Liver specific miRNAbinding sites can be engineered alone or further in combination withimmune cell (e.g., APC) miRNA binding sites in a polynucleotide of thedisclosure.

MiRNAs that are known to be expressed in the lung include, but are notlimited to, let-7a-2-3p, let-7a-3p, let-7a-5p, miR-126-3p, miR-126-5p,miR-127-3p, miR-127-5p, miR-130a-3p, miR-130a-5p, miR-130b-3p,miR-130b-5p, miR-133a, miR-133b, miR-134, miR-18a-3p, miR-18a-5p,miR-18b-3p, miR-18b-5p, miR-24-1-5p, miR-24-2-5p, miR-24-3p, miR-296-3p,miR-296-5p, miR-32-3p, miR-337-3p, miR-337-5p, miR-381-3p, andmiR-381-5p. MiRNA binding sites from any lung specific miRNA can beintroduced to or removed from a polynucleotide of the disclosure toregulate expression of the polynucleotide in the lung. Lung specificmiRNA binding sites can be engineered alone or further in combinationwith immune cell (e.g., APC) miRNA binding sites in a polynucleotide ofthe disclosure.

MiRNAs that are known to be expressed in the heart include, but are notlimited to, miR-1, miR-133a, miR-133b, miR-149-3p, miR-149-5p,miR-186-3p, miR-186-5p, miR-208a, miR-208b, miR-210, miR-296-3p,miR-320, miR-451a, miR-451b, miR-499a-3p, miR-499a-5p, miR-499b-3p,miR-499b-5p, miR-744-3p, miR-744-5p, miR-92b-3p, and miR-92b-5p. MiRNAbinding sites from any heart specific microRNA can be introduced to orremoved from a polynucleotide of the disclosure to regulate expressionof the polynucleotide in the heart. Heart specific miRNA binding sitescan be engineered alone or further in combination with immune cell(e.g., APC) miRNA binding sites in a polynucleotide of the disclosure.

MiRNAs that are known to be expressed in the nervous system include, butare not limited to, miR-124-5p, miR-125a-3p, miR-125a-5p, miR-125b-1-3p,miR-125b-2-3p, miR-125b-5p, miR-1271-3p, miR-1271-5p, miR-128,miR-132-5p, miR-135a-3p, miR-135a-5p, miR-135b-3p, miR-135b-5p, miR-137,miR-139-5p, miR-139-3p, miR-149-3p, miR-149-5p, miR-153, miR-181c-3p,miR-181c-5p, miR-183-3p, miR-183-5p, miR-190a, miR-190b, miR-212-3p,miR-212-5p, miR-219-1-3p, miR-219-2-3p, miR-23a-3p, miR-23a-5p,miR-30a-5p, miR-30b-3p, miR-30b-5p, miR-30c-1-3p, miR-30c-2-3p,miR-30c-5p, miR-30d-3p, miR-30d-5p, miR-329, miR-342-3p, miR-3665,miR-3666, miR 3p, miR-380-5p, miR-383, miR-410, miR-425-3p, miR-425-5p,miR-454-3p, miR-454-5p, miR-483, miR-510, miR-516a-3p, miR-548b-5p,miR-548c-5p, miR-571, miR-7-1-3p, miR-7-2-3p, miR-7-5p, miR-802,miR-922, miR-9-3p, and miR-9-5p. MiRNAs enriched in the nervous systemfurther include those specifically expressed in neurons, including, butnot limited to, miR-132-3p, miR-132-3p, miR-148b-3p, miR-148b-5p,miR-151a-3p, miR-151a-5p, miR-212-3p, miR-212-5p, miR-320b, miR-320e,miR-323a-3p, miR-323a-5p, miR-324-5p, miR-325, miR-326, miR-328, miR-922and those specifically expressed in glial cells, including, but notlimited to, miR-1250, miR-219-1-3p, miR-219-2-3p, miR-219-5p,miR-23a-3p, miR-23a-5p, miR-3065-3p, miR-3065-5p, miR-30e-3p,miR-30e-5p, miR-32-5p, miR-338-5p, and miR-657. MiRNA binding sites fromany CNS specific miRNA can be introduced to or removed from apolynucleotide of the disclosure to regulate expression of thepolynucleotide in the nervous system. Nervous system specific miRNAbinding sites can be engineered alone or further in combination withimmune cell (e.g., APC) miRNA binding sites in a polynucleotide of thedisclosure.

MiRNAs that are known to be expressed in the pancreas include, but arenot limited to, miR-105-3p, miR-105-5p, miR-184, miR-195-3p, miR-195-5p,miR-196a-3p, miR-196a-5p, miR-214-3p, miR-214-5p, miR-216a-3p,miR-216a-5p, miR-30a-3p, miR-33a-3p, miR-33a-5p, miR-375, miR-7-1-3p,miR-7-2-3p, miR-493-3p, miR-493-5p, and miR-944. MiRNA binding sitesfrom any pancreas specific miRNA can be introduced to or removed from apolynucleotide of the disclosure to regulate expression of thepolynucleotide in the pancreas. Pancreas specific miRNA binding sitescan be engineered alone or further in combination with immune cell (e.g.APC) miRNA binding sites in a polynucleotide of the disclosure.

MiRNAs that are known to be expressed in the kidney include, but are notlimited to, miR-122-3p, miR-145-5p, miR-17-5p, miR-192-3p, miR-192-5p,miR-194-3p, miR-194-5p, miR-20a-3p, miR-20a-5p, miR-204-3p, miR-204-5p,miR-210, miR-216a-3p, miR-216a-5p, miR-296-3p, miR-30a-3p, miR-30a-5p,miR-30b-3p, miR-30b-5p, miR-30c-1-3p, miR-30c-2-3p, miR30c-5p,miR-324-3p, miR-335-3p, miR-335-5p, miR-363-3p, miR-363-5p, and miR-562.MiRNA binding sites from any kidney specific miRNA can be introduced toor removed from a polynucleotide of the disclosure to regulateexpression of the polynucleotide in the kidney. Kidney specific miRNAbinding sites can be engineered alone or further in combination withimmune cell (e.g., APC) miRNA binding sites in a polynucleotide of thedisclosure.

MiRNAs that are known to be expressed in the muscle include, but are notlimited to, let-7g-3p, let-7g-5p, miR-1, miR-1286, miR-133a, miR-133b,miR-140-3p, miR-143-3p, miR-143-5p, miR-145-3p, miR-145-5p, miR-188-3p,miR-188-5p, miR-206, miR-208a, miR-208b, miR-25-3p, and miR-25-5p. MiRNAbinding sites from any muscle specific miRNA can be introduced to orremoved from a polynucleotide of the disclosure to regulate expressionof the polynucleotide in the muscle. Muscle specific miRNA binding sitescan be engineered alone or further in combination with immune cell(e.g., APC) miRNA binding sites in a polynucleotide of the disclosure.

MiRNAs are also differentially expressed in different types of cells,such as, but not limited to, endothelial cells, epithelial cells, andadipocytes.

MiRNAs that are known to be expressed in endothelial cells include, butare not limited to, let-7b-3p, let-7b-5p, miR-100-3p, miR-100-5p,miR-101-3p, miR-101-5p, miR-126-3p, miR-126-5p, miR-1236-3p,miR-1236-5p, miR-130a-3p, miR-130a-5p, miR-17-5p, miR-17-3p, miR-18a-3p,miR-18a-5p, miR-19a-3p, miR-19a-5p, miR-19b-1-5p, miR-19b-2-5p,miR-19b-3p, miR-20a-3p, miR-20a-5p, miR-217, miR-210, miR-21-3p,miR-21-5p, miR-221-3p, miR-221-5p, miR-222-3p, miR-222-5p, miR-23a-3p,miR-23a-5p, miR-296-5p, miR-361-3p, miR-361-5p, miR-421, miR-424-3p,miR-424-5p, miR-513a-5p, miR-92a-1-5p, miR-92a-2-5p, miR-92a-3p,miR-92b-3p, and miR-92b-5p. Many novel miRNAs are discovered inendothelial cells from deep-sequencing analysis (e.g., Voellenkle C etal., RNA, 2012, 18, 472-484, herein incorporated by reference in itsentirety). MiRNA binding sites from any endothelial cell specific miRNAcan be introduced to or removed from a polynucleotide of the disclosureto regulate expression of the polynucleotide in the endothelial cells.

MiRNAs that are known to be expressed in epithelial cells include, butare not limited to, let-7b-3p, let-7b-5p, miR-1246, miR-200a-3p,miR-200a-5p, miR-200b-3p, miR-200b-5p, miR-200c-3p, miR-200c-5p,miR-338-3p, miR-429, miR-451a, miR-451b, miR-494, miR-802 and miR-34a,miR-34b-5p, miR-34c-5p, miR-449a, miR-449b-3p, miR-449b-5p specific inrespiratory ciliated epithelial cells, let-7 family, miR-133a, miR-133b,miR-126 specific in lung epithelial cells, miR-382-3p, miR-382-5pspecific in renal epithelial cells, and miR-762 specific in cornealepithelial cells. MiRNA binding sites from any epithelial cell specificmiRNA can be introduced to or removed from a polynucleotide of thedisclosure to regulate expression of the polynucleotide in theepithelial cells.

In addition, a large group of miRNAs are enriched in embryonic stemcells, controlling stem cell self-renewal as well as the developmentand/or differentiation of various cell lineages, such as neural cells,cardiac, hematopoietic cells, skin cells, osteogenic cells and musclecells (e.g., Kuppusamy K T et al., Curr. Mol Med, 2013, 13(5), 757-764;Vidigal J A and Ventura A, Semin Cancer Biol. 2012, 22(5-6), 428-436;Goff L A et al., PLoS One, 2009, 4:e7192; Morin R D et al., Genome Res,2008,18, 610-621; Yoo J K et al., Stem Cells Dev. 2012, 21(11),2049-2057, each of which is herein incorporated by reference in itsentirety). MiRNAs abundant in embryonic stem cells include, but are notlimited to, let-7a-2-3p, let-a-3p, let-7a-5p, let7d-3p, let-7d-5p,miR-103a-2-3p, miR-103a-5p, miR-106b-3p, miR-106b-5p, miR-1246,miR-1275, miR-138-1-3p, miR-138-2-3p, miR-138-5p, miR-154-3p,miR-154-5p, miR-200c-3p, miR-200c-5p, miR-290, miR-301a-3p, miR-301a-5p,miR-302a-3p, miR-302a-5p, miR-302b-3p, miR-302b-5p, miR-302c-3p,miR-302c-5p, miR-302d-3p, miR-302d-5p, miR-302e, miR-367-3p, miR-367-5p,miR-369-3p, miR-369-5p, miR-370, miR-371, miR-373, miR-380-5p,miR-423-3p, miR-423-5p, miR-486-5p, miR-520c-3p, miR-548e, miR-548f,miR-548g-3p, miR-548g-5p, miR-548i, miR-548k, miR-5481, miR-548m,miR-548n, miR-5480-3p, miR-5480-5p, miR-548p, miR-664a-3p, miR-664a-5p,miR-664b-3p, miR-664b-5p, miR-766-3p, miR-766-5p, miR-885-3p,miR-885-5p, miR-93-3p, miR-93-5p, miR-941, miR-96-3p, miR-96-5p,miR-99b-3p and miR-99b-5p. Many predicted novel miRNAs are discovered bydeep sequencing in human embryonic stem cells (e.g., Morin R D et al.,Genome Res, 2008,18, 610-621; Goff L A et al., PLoS One, 2009, 4:e7192;Bar M et al., Stem cells, 2008, 26, 2496-2505, the content of each ofwhich is incorporated herein by reference in its entirety).

In one embodiment, the binding sites of embryonic stem cell specificmiRNAs can be included in or removed from the 3′UTR of a polynucleotideof the disclosure to modulate the development and/or differentiation ofembryonic stem cells, to inhibit the senescence of stem cells in adegenerative condition (e.g. degenerative diseases), or to stimulate thesenescence and apoptosis of stem cells in a disease condition (e.g.cancer stem cells).

MiRNA can also regulate complex biological processes such asangiogenesis (e.g., miR-132) (Anand and Cheresh Curr Opin Hematol 201118:171-176). In the polynucleotides of the disclosure, miRNA bindingsites that are involved in such processes can be removed or introduced,in order to tailor the expression of the polynucleotides to biologicallyrelevant cell types or relevant biological processes. In this context,the polynucleotides of the disclosure are defined as auxotrophicpolynucleotides.

In some embodiments, the microRNA binding site (e.g., miR-122 bindingsite) is fully complementary to miRNA (e.g., miR-122), thereby degradingthe mRNA fused to the miRNA binding site. In other embodiments, themiRNA binding site is not fully complementary to the correspondingmiRNA. In certain embodiments, the miRNA binding site (e.g., miR-122binding site) is the same length as the corresponding miRNA (e.g.,miR-122). In other embodiments, the microRNA binding site (e.g., miR-122binding site) is one nucleotide shorter than the corresponding microRNA(e.g., miR-122, which has 22 nts) at the 5′ terminus, the 3′ terminus,or both. In still other embodiments, the microRNA binding site (e.g.,miR-122 binding site) is two nucleotides shorter than the correspondingmicroRNA (e.g., miR-122) at the 5′ terminus, the 3′ terminus, or both.In yet other embodiments, the microRNA binding site (e.g., miR-122binding site) is three nucleotides shorter than the correspondingmicroRNA (e.g., miR-122) at the 5′ terminus, the 3′ terminus, or both.In some embodiments, the microRNA binding site (e.g., miR-122 bindingsite) is four nucleotides shorter than the corresponding microRNA (e.g.,miR-122) at the 5′ terminus, the 3′ terminus, or both. In otherembodiments, the microRNA binding site (e.g., miR-122 binding site) isfive nucleotides shorter than the corresponding microRNA (e.g., miR-122)at the 5′ terminus, the 3′ terminus, or both. In some embodiments, themicroRNA binding site (e.g., miR-122 binding site) is six nucleotidesshorter than the corresponding microRNA (e.g., miR-122) at the 5′terminus, the 3′ terminus, or both. In other embodiments, the microRNAbinding site (e.g., miR-122 binding site) is seven nucleotides shorterthan the corresponding microRNA (e.g., miR-122) at the 5′ terminus orthe 3′ terminus. In other embodiments, the microRNA binding site (e.g.,miR-122 binding site) is eight nucleotides shorter than thecorresponding microRNA (e.g., miR-122) at the 5′ terminus or the 3′terminus. In other embodiments, the microRNA binding site (e.g., miR-122binding site) is nine nucleotides shorter than the correspondingmicroRNA (e.g., miR-122) at the 5′ terminus or the 3′ terminus. In otherembodiments, the microRNA binding site (e.g., miR-122 binding site) isten nucleotides shorter than the corresponding microRNA (e.g., miR-122)at the 5′ terminus or the 3′ terminus. In other embodiments, themicroRNA binding site (e.g., miR-122 binding site) is eleven nucleotidesshorter than the corresponding microRNA (e.g., miR-122) at the 5′terminus or the 3′ terminus. In other embodiments, the microRNA bindingsite (e.g., miR-122 binding site) is twelve nucleotides shorter than thecorresponding microRNA (e.g., miR-122) at the 5′ terminus or the 3′terminus. The miRNA binding sites that are shorter than thecorresponding miRNAs are still capable of degrading the mRNAincorporating one or more of the miRNA binding sites or preventing themRNA from translation.

In some embodiments, the microRNA binding site (e.g., miR-122 bindingsite) has sufficient complementarity to miRNA (e.g., miR-122) so that aRISC complex comprising the miRNA (e.g., miR-122) cleaves thepolynucleotide comprising the microRNA binding site. In otherembodiments, the microRNA binding site (e.g., miR-122 binding site) hasimperfect complementarity so that a RISC complex comprising the miRNA(e.g., miR-122) induces instability in the polynucleotide comprising themicroRNA binding site. In another embodiment, the microRNA binding site(e.g., miR-122 binding site) has imperfect complementarity so that aRISC complex comprising the miRNA (e.g., miR-122) repressestranscription of the polynucleotide comprising the microRNA bindingsite. In one embodiment, the miRNA binding site (e.g., miR-122 bindingsite) has one mismatch from the corresponding miRNA (e.g., miR-122). Inanother embodiment, the miRNA binding site has two mismatches from thecorresponding miRNA. In other embodiments, the miRNA binding site hasthree mismatches from the corresponding miRNA. In other embodiments, themiRNA binding site has four mismatches from the corresponding miRNA. Insome embodiments, the miRNA binding site has five mismatches from thecorresponding miRNA. In other embodiments, the miRNA binding site hassix mismatches from the corresponding miRNA. In certain embodiments, themiRNA binding site has seven mismatches from the corresponding miRNA. Inother embodiments, the miRNA binding site has eight mismatches from thecorresponding miRNA. In other embodiments, the miRNA binding site hasnine mismatches from the corresponding miRNA. In other embodiments, themiRNA binding site has ten mismatches from the corresponding miRNA. Inother embodiments, the miRNA binding site has eleven mismatches from thecorresponding miRNA. In other embodiments, the miRNA binding site hastwelve mismatches from the corresponding miRNA.

In certain embodiments, the miRNA binding site (e.g., miR-122 bindingsite) has at least about ten contiguous nucleotides complementary to atleast about ten contiguous nucleotides of the corresponding miRNA (e.g.,miR-122), at least about eleven contiguous nucleotides complementary toat least about eleven contiguous nucleotides of the corresponding miRNA,at least about twelve contiguous nucleotides complementary to at leastabout twelve contiguous nucleotides of the corresponding miRNA, at leastabout thirteen contiguous nucleotides complementary to at least aboutthirteen contiguous nucleotides of the corresponding miRNA, or at leastabout fourteen contiguous nucleotides complementary to at least aboutfourteen contiguous nucleotides of the corresponding miRNA. In someembodiments, the miRNA binding sites have at least about fifteencontiguous nucleotides complementary to at least about fifteencontiguous nucleotides of the corresponding miRNA, at least aboutsixteen contiguous nucleotides complementary to at least about sixteencontiguous nucleotides of the corresponding miRNA, at least aboutseventeen contiguous nucleotides complementary to at least aboutseventeen contiguous nucleotides of the corresponding miRNA, at leastabout eighteen contiguous nucleotides complementary to at least abouteighteen contiguous nucleotides of the corresponding miRNA, at leastabout nineteen contiguous nucleotides complementary to at least aboutnineteen contiguous nucleotides of the corresponding miRNA, at leastabout twenty contiguous nucleotides complementary to at least abouttwenty contiguous nucleotides of the corresponding miRNA, or at leastabout twenty one contiguous nucleotides complementary to at least abouttwenty one contiguous nucleotides of the corresponding miRNA.

In some embodiments, the polynucleotides of the disclosure comprise atleast one miR122 binding site, at least two miR122 binding sites, atleast three miR122 binding sites, at least four miR122 binding sites, orat least five miR122 binding sites. In one aspect, the miRNA bindingsite binds miR-122 or is complementary to miR-122. In another aspect,the miRNA binding site binds to miR-122-3p or miR-122-5p. In aparticular aspect, the miRNA binding site comprises a nucleotidesequence at least 80%, at least 85%, at least 90%, at least 95%, or 100%identical to SEQ ID NO: 52 or 54, wherein the miRNA binding site bindsto miR-122. In another particular aspect, the miRNA binding sitecomprises a nucleotide sequence at least 80%, at least 85%, at least90%, at least 95%, or 100% identical to SEQ ID NO: 54, wherein the miRNAbinding site binds to miR-122 (e.g., with sufficient strength to induceRISC-mediated cleavage of the polynucleotide, e.g., mRNA, comprising themiRNA binding site). In another particular aspect, the miRNA bindingsite has less than 3 substitutions, less than 2 substitutions, or lessthan 1 substitution as compared to the miRNA binding site as set forthas SEQ ID NO: 54, wherein the miR binding site binds to miR-122 (e.g.,with sufficient strength to induce RISC-mediated cleavage of thepolynucleotide, e.g., mRNA, comprising the miRNA binding site).

In some embodiments, a miRNA binding site (e.g., miR-122 binding site)is inserted in the polynucleotide of the disclosure in any position ofthe polynucleotide (e.g., 3′ UTR); the insertion site in thepolynucleotide can be anywhere in the polynucleotide as long as theinsertion of the miRNA binding site in the polynucleotide does notinterfere with the translation of the functional IL-12 polypeptide orthe translation of the functional membrane domain in the absence of thecorresponding miRNA (e.g., miR122); and in the presence of the miRNA(e.g., miR122), the insertion of the miRNA binding site in thepolynucleotide and the binding of the miRNA binding site to thecorresponding miRNA are capable of degrading the polynucleotide orpreventing the translation of the polynucleotide. In one embodiment, amiRNA binding site is inserted in a 3′UTR of the polynucleotide.

In certain embodiments, a miRNA binding site is inserted in at leastabout 30 nucleotides downstream from a stop codon in a polynucleotide ofthe disclosure. In other embodiments, a miRNA binding site is insertedin at least about 10 nucleotides, at least about 15 nucleotides, atleast about 20 nucleotides, at least about 25 nucleotides, at leastabout 30 nucleotides, at least about 35 nucleotides, at least about 40nucleotides, at least about 45 nucleotides, at least about 50nucleotides, at least about 55 nucleotides, at least about 60nucleotides, at least about 65 nucleotides, at least about 70nucleotides, at least about 75 nucleotides, at least about 80nucleotides, at least about 85 nucleotides, at least about 90nucleotides, at least about 95 nucleotides, or at least about 100nucleotides downstream from a stop codon in a polynucleotide of thedisclosure. In other embodiments, a miRNA binding site is inserted inabout 10 nucleotides to about 100 nucleotides, about 20 nucleotides toabout 90 nucleotides, about 30 nucleotides to about 80 nucleotides,about 40 nucleotides to about 70 nucleotides, about 50 nucleotides toabout 60 nucleotides, about 45 nucleotides to about 65 nucleotidesdownstream from the stop codon in a polynucleotide of the disclosure. Insome embodiments, the miRNA binding site is inserted downstream of thestop codon in the nucleic acid sequence encoding an IL-12 polypeptide asdisclosed herein. In some embodiments, the miRNA binding site isinserted downstream of the stop codon in the nucleic acid sequenceencoding membrane domain as disclosed herein. In some embodiments, themiRNA binding site is inserted downstream of the stop codon in thenucleic acid sequence encoding heterologous polypeptide as disclosedherein

In some embodiments, a miRNA binding site is inserted in thepolynucleotide of the disclosure in any position of the polynucleotide(e.g., the 5′UTR and/or 3′UTR). In some embodiments, the 5′UTR comprisesa miRNA binding site. In some embodiments, the 3′UTR comprises a miRNAbinding site. In some embodiments, the 5′UTR and the 3′UTR comprise amiRNA binding site. The insertion site in the polynucleotide can beanywhere in the polynucleotide as long as the insertion of the miRNAbinding site in the polynucleotide does not interfere with thetranslation of a functional polypeptide in the absence of thecorresponding miRNA; and in the presence of the miRNA, the insertion ofthe miRNA binding site in the polynucleotide and the binding of themiRNA binding site to the corresponding miRNA are capable of degradingthe polynucleotide or preventing the translation of the polynucleotide.

In some embodiments, a miRNA binding site is inserted in at least about30 nucleotides downstream from the stop codon of an ORF in apolynucleotide of the disclosure comprising the ORF. In someembodiments, a miRNA binding site is inserted in at least about 10nucleotides, at least about 15 nucleotides, at least about 20nucleotides, at least about 25 nucleotides, at least about 30nucleotides, at least about 35 nucleotides, at least about 40nucleotides, at least about 45 nucleotides, at least about 50nucleotides, at least about 55 nucleotides, at least about 60nucleotides, at least about 65 nucleotides, at least about 70nucleotides, at least about 75 nucleotides, at least about 80nucleotides, at least about 85 nucleotides, at least about 90nucleotides, at least about 95 nucleotides, or at least about 100nucleotides downstream from the stop codon of an ORF in a polynucleotideof the disclosure. In some embodiments, a miRNA binding site is insertedin about 10 nucleotides to about 100 nucleotides, about 20 nucleotidesto about 90 nucleotides, about 30 nucleotides to about 80 nucleotides,about 40 nucleotides to about 70 nucleotides, about 50 nucleotides toabout 60 nucleotides, about 45 nucleotides to about 65 nucleotidesdownstream from the stop codon of an ORF in a polynucleotide of thedisclosure.

MiRNA gene regulation can be influenced by the sequence surrounding themiRNA such as, but not limited to, the species of the surroundingsequence, the type of sequence (e.g., heterologous, homologous,exogenous, endogenous, or artificial), regulatory elements in thesurrounding sequence and/or structural elements in the surroundingsequence. The miRNA can be influenced by the 5′UTR and/or 3′UTR. As anon-limiting example, a non-human 3′UTR can increase the regulatoryeffect of the miRNA sequence on the expression of a polypeptide ofinterest compared to a human 3′UTR of the same sequence type.

In one embodiment, other regulatory elements and/or structural elementsof the 5′UTR can influence miRNA mediated gene regulation. One exampleof a regulatory element and/or structural element is a structured IRES(Internal Ribosome Entry Site) in the 5′UTR, which is necessary for thebinding of translational elongation factors to initiate proteintranslation. EIF4A2 binding to this secondarily structured element inthe 5′-UTR is necessary for miRNA mediated gene expression (Meijer H Aet al., Science, 2013, 340, 82-85, herein incorporated by reference inits entirety). The polynucleotides of the disclosure can further includethis structured 5′UTR in order to enhance microRNA mediated generegulation.

At least one miRNA binding site can be engineered into the 3′UTR of apolynucleotide of the disclosure. In this context, at least two, atleast three, at least four, at least five, at least six, at least seven,at least eight, at least nine, at least ten, or more miRNA binding sitescan be engineered into a 3′UTR of a polynucleotide of the disclosure.For example, 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1to 3, 2, or 1 miRNA binding sites can be engineered into the 3′UTR of apolynucleotide of the disclosure. In one embodiment, miRNA binding sitesincorporated into a polynucleotide of the disclosure can be the same orcan be different miRNA sites. A combination of different miRNA bindingsites incorporated into a polynucleotide of the disclosure can includecombinations in which more than one copy of any of the different miRNAsites are incorporated. In another embodiment, miRNA binding sitesincorporated into a polynucleotide of the disclosure can target the sameor different tissues in the body. As a non-limiting example, through theintroduction of tissue-, cell-type-, or disease-specific miRNA bindingsites in the 3′-UTR of a polynucleotide of the disclosure, the degree ofexpression in specific cell types (e.g., hepatocytes, myeloid cells,endothelial cells, cancer cells, etc.) can be reduced.

In one embodiment, a miRNA binding site can be engineered near the 5′terminus of the 3′UTR, about halfway between the 5′ terminus and 3′terminus of the 3′UTR and/or near the 3′ terminus of the 3′UTR in apolynucleotide of the disclosure. As a non-limiting example, a miRNAbinding site can be engineered near the 5′ terminus of the 3′UTR andabout halfway between the 5′ terminus and 3′ terminus of the 3′UTR. Asanother non-limiting example, a miRNA binding site can be engineerednear the 3′ terminus of the 3′UTR and about halfway between the 5′terminus and 3′ terminus of the 3′UTR. As yet another non-limitingexample, a miRNA binding site can be engineered near the 5′ terminus ofthe 3′UTR and near the 3′ terminus of the 3′UTR.

In another embodiment, a 3′UTR can comprise 1, 2, 3, 4, 5, 6, 7, 8, 9,or 10 miRNA binding sites. The miRNA binding sites can be complementaryto a miRNA, miRNA seed sequence, and/or miRNA sequences flanking theseed sequence.

In one embodiment, a polynucleotide of the disclosure can be engineeredto include more than one miRNA site expressed in different tissues ordifferent cell types of a subject. As a non-limiting example, apolynucleotide of the disclosure can be engineered to include miR-192and miR-122 to regulate expression of the polynucleotide in the liverand kidneys of a subject. In another embodiment, a polynucleotide of thedisclosure can be engineered to include more than one miRNA site for thesame tissue.

In some embodiments, the therapeutic window and or differentialexpression associated with the polypeptide encoded by a polynucleotideof the disclosure can be altered with a miRNA binding site. For example,a polynucleotide encoding a polypeptide that provides a death signal canbe designed to be more highly expressed in cancer cells by virtue of themiRNA signature of those cells. Where a cancer cell expresses a lowerlevel of a particular miRNA, the polynucleotide encoding the bindingsite for that miRNA (or miRNAs) would be more highly expressed. Hence,the polypeptide that provides a death signal triggers or induces celldeath in the cancer cell. Neighboring non-cancer cells, harboring ahigher expression of the same miRNA would be less affected by theencoded death signal as the polynucleotide would be expressed at a lowerlevel due to the effects of the miRNA binding to the binding site or“sensor” encoded in the 3′UTR. Conversely, cell survival orcytoprotective signals can be delivered to tissues containing cancer andnon-cancerous cells where a miRNA has a higher expression in the cancercells—the result being a lower survival signal to the cancer cell and alarger survival signal to the normal cell. Multiple polynucleotides canbe designed and administered having different signals based on the useof miRNA binding sites as described herein.

In some embodiments, the expression of a polynucleotide of thedisclosure can be controlled by incorporating at least one sensorsequence in the polynucleotide and formulating the polynucleotide foradministration. As a non-limiting example, a polynucleotide of thedisclosure can be targeted to a tissue or cell by incorporating a miRNAbinding site and formulating the polynucleotide in a lipid nanoparticlecomprising a cationic lipid, including any of the lipids describedherein.

A polynucleotide of the disclosure can be engineered for more targetedexpression in specific tissues, cell types, or biological conditionsbased on the expression patterns of miRNAs in the different tissues,cell types, or biological conditions. Through introduction oftissue-specific miRNA binding sites, a polynucleotide of the disclosurecan be designed for optimal protein expression in a tissue or cell, orin the context of a biological condition.

In some embodiments, a polynucleotide of the disclosure can be designedto incorporate miRNA binding sites that either have 100% identity toknown miRNA seed sequences or have less than 100% identity to miRNA seedsequences. In some embodiments, a polynucleotide of the disclosure canbe designed to incorporate miRNA binding sites that have at least: 60%,65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity toknown miRNA seed sequences. The miRNA seed sequence can be partiallymutated to decrease miRNA binding affinity and as such result in reduceddown modulation of the polynucleotide. In essence, the degree of matchor mis-match between the miRNA binding site and the miRNA seed can actas a rheostat to more finely tune the ability of the miRNA to modulateprotein expression. In addition, mutation in the non-seed region of amiRNA binding site can also impact the ability of a miRNA to modulateprotein expression.

In one embodiment, a miRNA sequence can be incorporated into the loop ofa stem loop.

In another embodiment, a miRNA seed sequence can be incorporated in theloop of a stem loop and a miRNA binding site can be incorporated intothe 5′ or 3′ stem of the stem loop.

In one embodiment, a translation enhancer element (TEE) can beincorporated on the 5′end of the stem of a stem loop and a miRNA seedcan be incorporated into the stem of the stem loop. In anotherembodiment, a TEE can be incorporated on the 5′ end of the stem of astem loop, a miRNA seed can be incorporated into the stem of the stemloop and a miRNA binding site can be incorporated into the 3′ end of thestem or the sequence after the stem loop. The miRNA seed and the miRNAbinding site can be for the same and/or different miRNA sequences.

In one embodiment, the incorporation of a miRNA sequence and/or a TEEsequence changes the shape of the stem loop region which can increaseand/or decrease translation. (see e.g., Kedde et al., “A Pumilio-inducedRNA structure switch in p27-3′UTR controls miR-221 and miR-22accessibility.” Nature Cell Biology. 2010, incorporated herein byreference in its entirety).

In one embodiment, the 5′-UTR of a polynucleotide of the disclosure cancomprise at least one miRNA sequence. The miRNA sequence can be, but isnot limited to, a 19 or 22 nucleotide sequence and/or a miRNA sequencewithout the seed.

In one embodiment the miRNA sequence in the 5′UTR can be used tostabilize a polynucleotide of the disclosure described herein.

In another embodiment, a miRNA sequence in the 5′UTR of a polynucleotideof the disclosure can be used to decrease the accessibility of the siteof translation initiation such as, but not limited to a start codon.See, e.g., Matsuda et al., PLoS One. 2010 11(5):e15057; incorporatedherein by reference in its entirety, which used antisense locked nucleicacid (LNA) oligonucleotides and exon-junction complexes (EJCs) around astart codon (−4 to +37 where the A of the AUG codons is +1) in order todecrease the accessibility to the first start codon (AUG). Matsudashowed that altering the sequence around the start codon with an LNA orEJC affected the efficiency, length and structural stability of apolynucleotide. A polynucleotide of the disclosure can comprise a miRNAsequence, instead of the LNA or EJC sequence described by Matsuda et al,near the site of translation initiation in order to decrease theaccessibility to the site of translation initiation. The site oftranslation initiation can be prior to, after or within the miRNAsequence. As a non-limiting example, the site of translation initiationcan be located within a miRNA sequence such as a seed sequence orbinding site. As another non-limiting example, the site of translationinitiation may be located within a miR-122 sequence such as the seedsequence or the mir-122 binding site.

In some embodiments, a polynucleotide of the disclosure can include atleast one miRNA in order to dampen expression of the encoded polypeptidein a tissue or cell of interest. As a non-limiting example, apolynucleotide of the disclosure can include at least one miR-122binding site in order to dampen expression of an encoded polypeptide ofinterest in the liver. As another non-limiting example a polynucleotideof the disclosure can include at least one miR-142-3p binding site,miR-142-3p seed sequence, miR-142-3p binding site without the seed,miR-142-5p binding site, miR-142-5p seed sequence, miR-142-5p bindingsite without the seed, miR-146 binding site, miR-146 seed sequenceand/or miR-146 binding site without the seed sequence.

In some embodiments, a polynucleotide of the disclosure can comprise atleast one miRNA binding site in the 3′UTR in order to selectivelydegrade mRNA therapeutics in the immune cells to subdue unwantedimmunogenic reactions caused by therapeutic delivery. As a non-limitingexample, the miRNA binding site can make a polynucleotide of thedisclosure more unstable in antigen presenting cells. Non-limitingexamples of these miRNAs include mir-122-5p or mir-122-3p.

In one embodiment, a polynucleotide of the disclosure comprises at leastone miRNA sequence in a region of the polynucleotide that can interactwith a RNA binding protein.

In some embodiments, the polynucleotide of the disclosure (e.g., a RNA,e.g., an mRNA) comprises (i) a sequence-optimized nucleotide sequenceencoding an IL-12 polypeptide (e.g., the wild-type sequence, functionalfragment, or variant thereof), (ii) a sequence-optimized nucleotidesequence encoding a membrane domain, and (iii) a miRNA binding site(e.g., a miRNA binding site that binds to miR-122).

In some embodiments, the polynucleotide of the disclosure comprises anucleobase-modified sequence and a miRNA binding site disclosed herein,e.g., a miRNA binding site that binds to miR-122. In some embodiments,the polynucleotides of the disclosure comprising a miRNA binding siteare formulated with a delivery agent, e.g., a compound having theFormula (I).

16. 3′ UTR and the AU Rich Elements

In certain embodiments, a polynucleotide of the present disclosurefurther comprises a 3′ UTR. 3′-UTR is the section of mRNA thatimmediately follows the translation termination codon and often containsregulatory regions that post-transcriptionally influence geneexpression. Regulatory regions within the 3′-UTR can influencepolyadenylation, translation efficiency, localization, and stability ofthe mRNA. In one embodiment, the 3′-UTR useful for the disclosurecomprises a binding site for regulatory proteins or microRNAs.

In certain embodiments, the 3′ UTR useful for the polynucleotides of thedisclosure comprises a 3′UTR selected from those shown in thisapplication (e.g., SEQ ID NOs: 222-224). In some embodiments, the 3′UTRuseful for the polynucleotides of the disclosure comprises a 3′UTRcomprising the sequence set forth in SEQ ID NO: 283.

In certain embodiments, the 3′ UTR sequence useful for the disclosurecomprises a nucleotide sequence at least about 60%, at least about 70%,at least about 80%, at least about 90%, at least about 95%, at leastabout 96%, at least about 97%, at least about 98%, at least about 99%,or about 100% identical to a sequence selected from the group consistingof 3′UTR sequences listed herein and any combination thereof.

17. Regions Having a 5′ Cap

The disclosure also includes a polynucleotide that comprises both a 5′Cap and a polynucleotide of the present disclosure.

The 5′ cap structure of a natural mRNA is involved in nuclear export,increasing mRNA stability and binds the mRNA Cap Binding Protein (CBP),which is responsible for mRNA stability in the cell and translationcompetency through the association of CBP with poly(A) binding proteinto form the mature cyclic mRNA species. The cap further assists theremoval of 5′ proximal introns during mRNA splicing.

Endogenous mRNA molecules can be 5′-end capped generating a5′-ppp-5′-triphosphate linkage between a terminal guanosine cap residueand the 5′-terminal transcribed sense nucleotide of the mRNA molecule.This 5′-guanylate cap can then be methylated to generate anN7-methyl-guanylate residue. The ribose sugars of the terminal and/oranteterminal transcribed nucleotides of the 5′ end of the mRNA canoptionally also be 2′-O-methylated. 5′-decapping through hydrolysis andcleavage of the guanylate cap structure can target a nucleic acidmolecule, such as an mRNA molecule, for degradation.

In some embodiments, the polynucleotides of the present disclosureincorporate a cap moiety.

In some embodiments, polynucleotides of the present disclosure comprisea non-hydrolyzable cap structure preventing decapping and thusincreasing mRNA half-life. Because cap structure hydrolysis requirescleavage of 5′-ppp-5′ phosphorodiester linkages, modified nucleotidescan be used during the capping reaction. For example, a Vaccinia CappingEnzyme from New England Biolabs (Ipswich, Mass.) can be used withα-thio-guanosine nucleotides according to the manufacturer'sinstructions to create a phosphorothioate linkage in the 5′-ppp-5′ cap.Additional modified guanosine nucleotides can be used such asα-methyl-phosphonate and seleno-phosphate nucleotides.

Additional modifications include, but are not limited to,2′-O-methylation of the ribose sugars of 5′-terminal and/or5′-anteterminal nucleotides of the polynucleotide (as mentioned above)on the 2′-hydroxyl group of the sugar ring. Multiple distinct 5′-capstructures can be used to generate the 5′-cap of a nucleic acidmolecule, such as a polynucleotide that functions as an mRNA molecule.Cap analogs, which herein are also referred to as synthetic cap analogs,chemical caps, chemical cap analogs, or structural or functional capanalogs, differ from natural (i.e., endogenous, wild-type orphysiological) 5′-caps in their chemical structure, while retaining capfunction. Cap analogs can be chemically (i.e., non-enzymatically) orenzymatically synthesized and/or linked to the polynucleotides of thedisclosure.

For example, the Anti-Reverse Cap Analog (ARCA) cap contains twoguanines linked by a 5′-5′-triphosphate group, wherein one guaninecontains an N7 methyl group as well as a 3′-O-methyl group (i.e.,N7,3′-O-dimethyl-guanosine-5′-triphosphate-5′-guanosine (m⁷G-3′mppp-G;which may equivalently be designated 3′O-Me-m7G(5)ppp(5′)G). The 3′-0atom of the other, unmodified, guanine becomes linked to the 5′-terminalnucleotide of the capped polynucleotide. The N7- and 3′-O-methlyatedguanine provides the terminal moiety of the capped polynucleotide.Another exemplary cap is mCAP, which is similar to ARCA but has a2′-O-methyl group on guanosine (i.e.,N7,2′-O-dimethyl-guanosine-5¹-triphosphate-5′-guanosine, m⁷Gm-ppp-G).

In some embodiments, the cap is a dinucleotide cap analog. As anon-limiting example, the dinucleotide cap analog can be modified atdifferent phosphate positions with a boranophosphate group or aphophoroselenoate group such as the dinucleotide cap analogs describedin U.S. Pat. No. 8,519,110, the contents of which are hereinincorporated by reference in its entirety.

In another embodiment, the cap is a cap analog is aN7-(4-chlorophenoxyethyl) substituted dinucleotide form of a cap analogknown in the art and/or described herein. Non-limiting examples of aN7-(4-chlorophenoxyethyl) substituted dinucleotide form of a cap analoginclude a N7-(4-chlorophenoxyethyl)-G(5′)ppp(5′)G and aN7-(4-chlorophenoxyethyl)-m^(3′-O)G(5)ppp(5′)G cap analog (See, e.g.,the various cap analogs and the methods of synthesizing cap analogsdescribed in Kore et al. Bioorganic & Medicinal Chemistry 201321:4570-4574; the contents of which are herein incorporated by referencein its entirety). In another embodiment, a cap analog of the presentdisclosure is a 4-chloro/bromophenoxyethyl analog.

While cap analogs allow for the concomitant capping of a polynucleotideor a region thereof, in an in vitro transcription reaction, up to 20% oftranscripts can remain uncapped. This, as well as the structuraldifferences of a cap analog from an endogenous 5′-cap structures ofnucleic acids produced by the endogenous, cellular transcriptionmachinery, can lead to reduced translational competency and reducedcellular stability.

Polynucleotides of the disclosure can also be capped post-manufacture(whether IVT or chemical synthesis), using enzymes, in order to generatemore authentic 5′-cap structures. As used herein, the phrase “moreauthentic” refers to a feature that closely mirrors or mimics, eitherstructurally or functionally, an endogenous or wild type feature. Thatis, a “more authentic” feature is better representative of anendogenous, wild-type, natural or physiological cellular function and/orstructure as compared to synthetic features or analogs, etc., of theprior art, or which outperforms the corresponding endogenous, wild-type,natural or physiological feature in one or more respects. Non-limitingexamples of more authentic 5′cap structures of the present disclosureare those that, among other things, have enhanced binding of cap bindingproteins, increased half-life, reduced susceptibility to 5′endonucleases and/or reduced 5′decapping, as compared to synthetic 5′capstructures known in the art (or to a wild-type, natural or physiological5′cap structure). For example, recombinant Vaccinia Virus Capping Enzymeand recombinant 2′-O-methyltransferase enzyme can create a canonical5′-5′-triphosphate linkage between the 5′-terminal nucleotide of apolynucleotide and a guanine cap nucleotide wherein the cap guaninecontains an N7 methylation and the 5′-terminal nucleotide of the mRNAcontains a 2′-O-methyl. Such a structure is termed the Cap 1 structure.This cap results in a higher translational-competency and cellularstability and a reduced activation of cellular pro-inflammatorycytokines, as compared, e.g., to other 5′cap analog structures known inthe art. Cap structures include, but are not limited to,7mG(5′)ppp(5′)N,pN2p (cap 0), 7mG(5′)ppp(5′)NlmpNp (cap 1), and7mG(5′)-ppp(5′)NlmpN2mp (cap 2).

As a non-limiting example, capping chimeric polynucleotidespost-manufacture can be more efficient as nearly 100% of the chimericpolynucleotides can be capped. This is in contrast to ˜80% when a capanalog is linked to a chimeric polynucleotide in the course of an invitro transcription reaction.

According to the present disclosure, 5′ terminal caps can includeendogenous caps or cap analogs. According to the present disclosure, a5′ terminal cap can comprise a guanine analog. Useful guanine analogsinclude, but are not limited to, inosine, N1-methyl-guanosine,2′fluoro-guanosine, 7-deaza-guanosine, 8-oxo-guanosine,2-amino-guanosine, LNA-guanosine, and 2-azido-guanosine.

18. Poly-A Tails

In some embodiments, the polynucleotides of the present disclosurefurther comprise a poly-A tail. In further embodiments, terminal groupson the poly-A tail can be incorporated for stabilization. In otherembodiments, a poly-A tail comprises des-3′ hydroxyl tails.

During RNA processing, a long chain of adenine nucleotides (poly-A tail)can be added to a polynucleotide such as an mRNA molecule in order toincrease stability. Immediately after transcription, the 3′ end of thetranscript can be cleaved to free a 3′ hydroxyl. Then poly-A polymeraseadds a chain of adenine nucleotides to the RNA. The process, calledpolyadenylation, adds a poly-A tail that can be between, for example,approximately 80 to approximately 250 residues long, includingapproximately 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190,200, 210, 220, 230, 240 or 250 residues long.

PolyA tails can also be added after the construct is exported from thenucleus.

According to the present disclosure, terminal groups on the polyA tailcan be incorporated for stabilization. Polynucleotides of the presentdisclosure can include des-3′ hydroxyl tails. They can also includestructural moieties or 2′-Omethyl modifications as taught by Junjie Li,et al. (Current Biology, Vol. 15, 1501-1507, Aug. 23, 2005, the contentsof which are incorporated herein by reference in its entirety).

The polynucleotides of the present disclosure can be designed to encodetranscripts with alternative polyA tail structures including histonemRNA. According to Norbury, “Terminal uridylation has also been detectedon human replication-dependent histone mRNAs. The turnover of thesemRNAs is thought to be important for the prevention of potentially toxichistone accumulation following the completion or inhibition ofchromosomal DNA replication. These mRNAs are distinguished by their lackof a 3′ poly(A) tail, the function of which is instead assumed by astable stem—loop structure and its cognate stem—loop binding protein(SLBP); the latter carries out the same functions as those of PABP onpolyadenylated mRNAs” (Norbury, “Cytoplasmic RNA: a case of the tailwagging the dog,” Nature Reviews Molecular Cell Biology; AOP, publishedonline 29 Aug. 2013; doi:10.1038/nrm3645) the contents of which areincorporated herein by reference in its entirety.

Unique poly-A tail lengths provide certain advantages to thepolynucleotides of the present disclosure. Generally, the length of apoly-A tail, when present, is greater than 30 nucleotides in length. Inanother embodiment, the poly-A tail is greater than 35 nucleotides inlength (e.g., at least or greater than about 35, 40, 45, 50, 55, 60, 70,80, 90, 100, 120, 140, 160, 180, 200, 250, 300, 350, 400, 450, 500, 600,700, 800, 900, 1,000, 1,100, 1,200, 1,300, 1,400, 1,500, 1,600, 1,700,1,800, 1,900, 2,000, 2,500, and 3,000 nucleotides).

In some embodiments, the polynucleotide or region thereof includes fromabout 30 to about 3,000 nucleotides (e.g., from 30 to 50, from 30 to100, from 30 to 250, from 30 to 500, from 30 to 750, from 30 to 1,000,from 30 to 1,500, from 30 to 2,000, from 30 to 2,500, from 50 to 100,from 50 to 250, from 50 to 500, from 50 to 750, from 50 to 1,000, from50 to 1,500, from 50 to 2,000, from 50 to 2,500, from 50 to 3,000, from100 to 500, from 100 to 750, from 100 to 1,000, from 100 to 1,500, from100 to 2,000, from 100 to 2,500, from 100 to 3,000, from 500 to 750,from 500 to 1,000, from 500 to 1,500, from 500 to 2,000, from 500 to2,500, from 500 to 3,000, from 1,000 to 1,500, from 1,000 to 2,000, from1,000 to 2,500, from 1,000 to 3,000, from 1,500 to 2,000, from 1,500 to2,500, from 1,500 to 3,000, from 2,000 to 3,000, from 2,000 to 2,500,and from 2,500 to 3,000).

In some embodiments, the poly-A tail is designed relative to the lengthof the overall polynucleotide or the length of a particular region ofthe polynucleotide. This design can be based on the length of a codingregion, the length of a particular feature or region or based on thelength of the ultimate product expressed from the polynucleotides.

In this context, the poly-A tail can be 10, 20, 30, 40, 50, 60, 70, 80,90, or 100% greater in length than the polynucleotide or featurethereof. The poly-A tail can also be designed as a fraction of thepolynucleotides to which it belongs. In this context, the poly-A tailcan be 10, 20, 30, 40, 50, 60, 70, 80, or 90% or more of the totallength of the construct, a construct region or the total length of theconstruct minus the poly-A tail. Further, engineered binding sites andconjugation of polynucleotides for Poly-A binding protein can enhanceexpression.

Additionally, multiple distinct polynucleotides can be linked togethervia the PABP (Poly-A binding protein) through the 3′-end using modifiednucleotides at the 3′-terminus of the poly-A tail. Transfectionexperiments can be conducted in relevant cell lines at and proteinproduction can be assayed by ELISA at 12 hr, 24 hr, 48 hr, 72 hr and day7 post-transfection.

In some embodiments, the polynucleotides of the present disclosure aredesigned to include a polyA-G Quartet region. The G-quartet is a cyclichydrogen bonded array of four guanine nucleotides that can be formed byG-rich sequences in both DNA and RNA. In this embodiment, the G-quartetis incorporated at the end of the poly-A tail. The resultantpolynucleotide is assayed for stability, protein production and otherparameters including half-life at various time points. It has beendiscovered that the polyA-G quartet results in protein production froman mRNA equivalent to at least 75% of that seen using a poly-A tail of120 nucleotides alone.

19. Start Codon Region

The disclosure also includes a polynucleotide that comprises both astart codon region and the polynucleotide described herein. In someembodiments, the polynucleotides of the present disclosure can haveregions that are analogous to or function like a start codon region.

In some embodiments, the translation of a polynucleotide can initiate ona codon that is not the start codon AUG. Translation of thepolynucleotide can initiate on an alternative start codon such as, butnot limited to, ACG, AGG, AAG, CTG/CUG, GTG/GUG, ATA/AUA, ATT/AUU,TTG/UUG (see Touriol et al. Biology of the Cell 95 (2003) 169-178 andMatsuda and Mauro PLoS ONE, 2010 5:11; the contents of each of which areherein incorporated by reference in its entirety).

As a non-limiting example, the translation of a polynucleotide begins onthe alternative start codon ACG. As another non-limiting example,polynucleotide translation begins on the alternative start codon CTG orCUG. As yet another non-limiting example, the translation of apolynucleotide begins on the alternative start codon GTG or GUG.

Nucleotides flanking a codon that initiates translation such as, but notlimited to, a start codon or an alternative start codon, are known toaffect the translation efficiency, the length and/or the structure ofthe polynucleotide. (See, e.g., Matsuda and Mauro PLoS ONE, 2010 5:11;the contents of which are herein incorporated by reference in itsentirety). Masking any of the nucleotides flanking a codon thatinitiates translation can be used to alter the position of translationinitiation, translation efficiency, length and/or structure of apolynucleotide.

In some embodiments, a masking agent can be used near the start codon oralternative start codon in order to mask or hide the codon to reduce theprobability of translation initiation at the masked start codon oralternative start codon. Non-limiting examples of masking agents includeantisense locked nucleic acids (LNA) polynucleotides and exon-junctioncomplexes (EJCs) (See, e.g., Matsuda and Mauro describing masking agentsLNA polynucleotides and EJCs (PLoS ONE, 2010 5:11); the contents ofwhich are herein incorporated by reference in its entirety).

In another embodiment, a masking agent can be used to mask a start codonof a polynucleotide in order to increase the likelihood that translationwill initiate on an alternative start codon. In some embodiments, amasking agent can be used to mask a first start codon or alternativestart codon in order to increase the chance that translation willinitiate on a start codon or alternative start codon downstream to themasked start codon or alternative start codon.

In some embodiments, a start codon or alternative start codon can belocated within a perfect complement for a miR binding site. The perfectcomplement of a miR binding site can help control the translation,length and/or structure of the polynucleotide similar to a maskingagent. As a non-limiting example, the start codon or alternative startcodon can be located in the middle of a perfect complement for a miRNAbinding site. The start codon or alternative start codon can be locatedafter the first nucleotide, second nucleotide, third nucleotide, fourthnucleotide, fifth nucleotide, sixth nucleotide, seventh nucleotide,eighth nucleotide, ninth nucleotide, tenth nucleotide, eleventhnucleotide, twelfth nucleotide, thirteenth nucleotide, fourteenthnucleotide, fifteenth nucleotide, sixteenth nucleotide, seventeenthnucleotide, eighteenth nucleotide, nineteenth nucleotide, twentiethnucleotide or twenty-first nucleotide.

In another embodiment, the start codon of a polynucleotide can beremoved from the polynucleotide sequence in order to have thetranslation of the polynucleotide begin on a codon that is not the startcodon. Translation of the polynucleotide can begin on the codonfollowing the removed start codon or on a downstream start codon or analternative start codon. In a non-limiting example, the start codon ATGor AUG is removed as the first 3 nucleotides of the polynucleotidesequence in order to have translation initiate on a downstream startcodon or alternative start codon. The polynucleotide sequence where thestart codon was removed can further comprise at least one masking agentfor the downstream start codon and/or alternative start codons in orderto control or attempt to control the initiation of translation, thelength of the polynucleotide and/or the structure of the polynucleotide.

20. Stop Codon Region

The disclosure also includes a polynucleotide that comprises both a stopcodon region and the nucleic acid sequences described herein. In someembodiments, the polynucleotides of the present disclosure can includeat least two stop codons before the 3′ untranslated region (UTR). Thestop codon can be selected from TGA, TAA and TAG in the case of DNA, orfrom UGA, UAA and UAG in the case of RNA. In some embodiments, thepolynucleotides of the present disclosure include the stop codon TGA inthe case or DNA, or the stop codon UGA in the case of RNA, and oneadditional stop codon. In a further embodiment the addition stop codoncan be TAA or UAA. In another embodiment, the polynucleotides of thepresent disclosure include three consecutive stop codons, four stopcodons, or more.

21. Insertions and Substitutions

The disclosure also includes a polynucleotide of the present disclosurethat further comprises insertions and/or substitutions.

In some embodiments, the 5′UTR of the polynucleotide can be replaced bythe insertion of at least one region and/or string of nucleosides of thesame base. The region and/or string of nucleotides can include, but isnot limited to, at least 3, at least 4, at least 5, at least 6, at least7 or at least 8 nucleotides and the nucleotides can be natural and/orunnatural. As a non-limiting example, the group of nucleotides caninclude 5-8 adenine, cytosine, thymine, a string of any of the othernucleotides disclosed herein and/or combinations thereof.

In some embodiments, the 5′UTR of the polynucleotide can be replaced bythe insertion of at least two regions and/or strings of nucleotides oftwo different bases such as, but not limited to, adenine, cytosine,thymine, any of the other nucleotides disclosed herein and/orcombinations thereof. For example, the 5′UTR can be replaced byinserting 5-8 adenine bases followed by the insertion of 5-8 cytosinebases. In another example, the 5′UTR can be replaced by inserting 5-8cytosine bases followed by the insertion of 5-8 adenine bases.

In some embodiments, the polynucleotide can include at least onesubstitution and/or insertion downstream of the transcription start sitethat can be recognized by an RNA polymerase. As a non-limiting example,at least one substitution and/or insertion can occur downstream of thetranscription start site by substituting at least one nucleic acid inthe region just downstream of the transcription start site (such as, butnot limited to, +1 to +6). Changes to region of nucleotides justdownstream of the transcription start site can affect initiation rates,increase apparent nucleotide triphosphate (NTP) reaction constantvalues, and increase the dissociation of short transcripts from thetranscription complex curing initial transcription (Brieba et al,Biochemistry (2002) 41: 5144-5149; herein incorporated by reference inits entirety). The modification, substitution and/or insertion of atleast one nucleoside can cause a silent mutation of the sequence or cancause a mutation in the amino acid sequence.

In some embodiments, the polynucleotide can include the substitution ofat least 1, at least 2, at least 3, at least 4, at least 5, at least 6,at least 7, at least 8, at least 9, at least 10, at least 11, at least12 or at least 13 guanine bases downstream of the transcription startsite.

In some embodiments, the polynucleotide can include the substitution ofat least 1, at least 2, at least 3, at least 4, at least 5 or at least 6guanine bases in the region just downstream of the transcription startsite. As a non-limiting example, if the nucleotides in the region areGGGAGA, the guanine bases can be substituted by at least 1, at least 2,at least 3 or at least 4 adenine nucleotides. In another non-limitingexample, if the nucleotides in the region are GGGAGA the guanine basescan be substituted by at least 1, at least 2, at least 3 or at least 4cytosine bases. In another non-limiting example, if the nucleotides inthe region are GGGAGA the guanine bases can be substituted by at least1, at least 2, at least 3 or at least 4 thymine, and/or any of thenucleotides described herein.

In some embodiments, the polynucleotide can include at least onesubstitution and/or insertion upstream of the start codon. For thepurpose of clarity, one of skill in the art would appreciate that thestart codon is the first codon of the protein coding region whereas thetranscription start site is the site where transcription begins. Thepolynucleotide can include, but is not limited to, at least 1, at least2, at least 3, at least 4, at least 5, at least 6, at least 7 or atleast 8 substitutions and/or insertions of nucleotide bases. Thenucleotide bases can be inserted or substituted at 1, at least 1, atleast 2, at least 3, at least 4 or at least 5 locations upstream of thestart codon. The nucleotides inserted and/or substituted can be the samebase (e.g., all A or all C or all T or all G), two different bases(e.g., A and C, A and T, or C and T), three different bases (e.g., A, Cand T or A, C and T) or at least four different bases.

As a non-limiting example, the guanine base upstream of the codingregion in the polynucleotide can be substituted with adenine, cytosine,thymine, or any of the nucleotides described herein. In anothernon-limiting example the substitution of guanine bases in thepolynucleotide can be designed so as to leave one guanine base in theregion downstream of the transcription start site and before the startcodon (see Esvelt et al. Nature (2011) 472(7344):499-503; the contentsof which is herein incorporated by reference in its entirety). As anon-limiting example, at least 5 nucleotides can be inserted at 1location downstream of the transcription start site but upstream of thestart codon and the at least 5 nucleotides can be the same base type.

22. Polynucleotide Comprising an mRNA Encoding a Tethered IL-12Polypeptide

In certain embodiments, a polynucleotide of the present disclosure, forexample a polynucleotide comprising an mRNA nucleotide sequence encodinga tethered IL-12 polypeptide comprises from 5′ to 3′ end:

-   -   (i) a 5′ UTR, such as the sequences provided above, comprising a        5′ cap provided above;    -   (ii) a nucleic acid sequence encoding an IL-12 polypeptide        disclosed herein, e.g., a nucleic acid sequence encoding IL-12B,        a nucleic acid sequence encoding IL-12A, and, optionally, a        nucleic acid sequence encoding a linker as disclosed herein that        connects IL-12B and IL-12A, e.g., a sequence optimized nucleic        acid sequence encoding an IL-12 polypeptide disclosed herein;    -   (iii) optionally, a nucleic acid sequence encoding a linker as        disclosed herein that connects the IL-12 polypeptide to a        membrane domain as disclosed herein;    -   (iv) a nucleic acid sequence encoding a membrane domain as        disclosed herein, e.g., a transmembrane domain as disclosed        herein, e.g., a Type I transmembrane domain, e.g., a CD8, CD80,        or PDGF-R transmembrane domain;    -   (v) at least one stop codon;    -   (vi) a 3′ UTR, such as the sequences provided above; and    -   (vii) a poly-A tail provided above.

In certain embodiments, a polynucleotide of the present disclosure, forexample a polynucleotide comprising an mRNA nucleotide sequence encodinga tethered IL-12 polypeptide comprises from 5′ to 3′ end:

-   -   (i) a 5′ UTR, such as the sequences provided above, comprising a        5′ cap;    -   (ii) a nucleic acid sequence encoding a membrane domain as        disclosed herein, e.g., a transmembrane domain as disclosed        herein, e.g., a Type II transmembrane domain;    -   (iii) optionally, a nucleic acid sequence encoding a linker as        disclosed herein that connects the membrane domain as disclosed        herein to an IL-12 polypeptide as disclosed herein;    -   (iv) a nucleic acid sequence encoding an IL-12 polypeptide        disclosed herein, e.g., a nucleic acid sequence encoding IL-12B,        a nucleic acid sequence encoding IL-12A, and, optionally, a        nucleic acid sequence encoding a linker as disclosed herein that        connects IL-12B and IL-12A, e.g., a sequence optimized nucleic        acid sequence encoding an IL-12 polypeptide disclosed herein;    -   (v) at least one stop codon;    -   (vi) a 3′ UTR, such as the sequences provided above; and    -   (vii) a poly-A tail provided above.

In some embodiments, the polynucleotide further comprises a miRNAbinding site, e.g., a miRNA binding site that binds to miRNA-122. Insome embodiments, the 5′UTR comprises the miRNA binding site.

In some embodiments, a polynucleotide of the present disclosurecomprises a nucleotide sequence encoding a polypeptide sequence at least70%, at least 80%, at least 81%, at least 82%, at least 83%, at least84%, at least 85%, at least 86%, at least 87%, at least 88%, at least89%, at least 90%, at least 91%, at least 92%, at least 93%, at least94%, at least 95%, at least 96%, at least 97%, at least 98%, at least99%, or 100% identical to the protein sequence of a wild type IL-12(e.g., isoform 1, 2, 3, or 4).

23. Methods of Making Polynucleotides

The present disclosure also provides methods for making a polynucleotideof the disclosure or a complement thereof.

In some aspects, a polynucleotide (e.g., a RNA, e.g., an mRNA) disclosedherein can be constructed using in vitro transcription. In otheraspects, a polynucleotide (e.g., a RNA, e.g., an mRNA) disclosed hereincan be constructed by chemical synthesis using an oligonucleotidesynthesizer.

In other aspects, a polynucleotide (e.g., a RNA, e.g., an mRNA)disclosed herein is made by using a host cell. In certain aspects, apolynucleotide (e.g., a RNA, e.g., an mRNA) disclosed herein is made byone or more combination of the IVT, chemical synthesis, host cellexpression, or any other methods known in the art. In other embodiments,a host cell is a eukaryotic cell, e.g., in vitro mammalian cells.

Naturally occurring nucleosides, non-naturally occurring nucleosides, orcombinations thereof, can totally or partially naturally replaceoccurring nucleosides present in the candidate nucleotide sequence andcan be incorporated into a sequence-optimized nucleotide sequence (e.g.,a RNA, e.g., an mRNA) as disclosed herein. The resultantpolynucleotides, e.g., mRNAs, can then be examined for their ability toproduce protein and/or produce a therapeutic outcome.

a. In Vitro Transcription/Enzymatic Synthesis

The polynucleotides of the present disclosure disclosed herein can betranscribed using an in vitro transcription (IVT) system. The systemtypically comprises a transcription buffer, nucleotide triphosphates(NTPs), an RNase inhibitor and a polymerase. The NTPs can be selectedfrom, but are not limited to, those described herein including naturaland unnatural (modified) NTPs. The polymerase can be selected from, butis not limited to, T7 RNA polymerase, T3 RNA polymerase and mutantpolymerases such as, but not limited to, polymerases able to incorporatepolynucleotides disclosed herein. See U.S. Publ. No. US20130259923,which is herein incorporated by reference in its entirety.

Any number of RNA polymerases or variants can be used in the synthesisof the polynucleotides of the present disclosure. RNA polymerases can bemodified by inserting or deleting amino acids of the RNA polymerasesequence. As a non-limiting example, the RNA polymerase can be modifiedto exhibit an increased ability to incorporate a 2′-modified nucleotidetriphosphate compared to an unmodified RNA polymerase (see InternationalPublication WO2008078180 and U.S. Pat. No. 8,101,385; hereinincorporated by reference in their entireties).

Variants can be obtained by evolving an RNA polymerase, optimizing theRNA polymerase amino acid and/or nucleic acid sequence and/or by usingother methods known in the art. As a non-limiting example, T7 RNApolymerase variants can be evolved using the continuous directedevolution system set out by Esvelt et al. (Nature 472:499-503 (2011);herein incorporated by reference in its entirety) where clones of T7 RNApolymerase can encode at least one mutation such as, but not limited to,lysine at position 93 substituted for threonine (K93T), I4M, A7T, E63V,V64D, A65E, D66Y, T76N, C125R, S128R, A136T, N165S, G175R, H176L, Y178H,F182L, L196F, G198V, D208Y, E222K, S228A, Q239R, T243N, G259D, M2671,G280C, H300R, D351A, A354S, E356D, L360P, A383V, Y385C, D388Y, S397R,M401T, N410S, K450R, P451T, G452V, E484A, H523L, H524N, G542V, E565K,K577E, K577M, N601S, S684Y, L6991, K713E, N748D, Q754R, E775K, A827V,D851N or L864F. As another non-limiting example, T7 RNA polymerasevariants can encode at least mutation as described in U.S. Pub. Nos.20100120024 and 20070117112; herein incorporated by reference in theirentireties. Variants of RNA polymerase can also include, but are notlimited to, substitutional variants, conservative amino acidsubstitution, insertional variants, deletional variants and/or covalentderivatives.

In one aspect, the polynucleotide can be designed to be recognized bythe wild type or variant RNA polymerases. In doing so, thepolynucleotide can be modified to contain sites or regions of sequencechanges from the wild type or parent chimeric polynucleotide.

Polynucleotide or nucleic acid synthesis reactions can be carried out byenzymatic methods utilizing polymerases. Polymerases catalyze thecreation of phosphodiester bonds between nucleotides in a polynucleotideor nucleic acid chain. Currently known DNA polymerases can be dividedinto different families based on amino acid sequence comparison andcrystal structure analysis. DNA polymerase I (pol I) or A polymerasefamily, including the Klenow fragments of E. coli, Bacillus DNApolymerase I, Thermus aquaticus (Taq) DNA polymerases, and the T7 RNAand DNA polymerases, is among the best studied of these families.Another large family is DNA polymerase a (pol a) or B polymerase family,including all eukaryotic replicating DNA polymerases and polymerasesfrom phages T4 and RB69. Although they employ similar catalyticmechanism, these families of polymerases differ in substratespecificity, substrate analog-incorporating efficiency, degree and ratefor primer extension, mode of DNA synthesis, exonuclease activity, andsensitivity against inhibitors.

DNA polymerases are also selected based on the optimum reactionconditions they require, such as reaction temperature, pH, and templateand primer concentrations. Sometimes a combination of more than one DNApolymerases is employed to achieve the desired DNA fragment size andsynthesis efficiency. For example, Cheng et al. increase pH, addglycerol and dimethyl sulfoxide, decrease denaturation times, increaseextension times, and utilize a secondary thermostable DNA polymerasethat possesses a 3′ to 5′ exonuclease activity to effectively amplifylong targets from cloned inserts and human genomic DNA. (Cheng et al.,PNAS 91:5695-5699 (1994), the contents of which are incorporated hereinby reference in their entirety). RNA polymerases from bacteriophage T3,T7, and SP6 have been widely used to prepare RNAs for biochemical andbiophysical studies. RNA polymerases, capping enzymes, and poly-Apolymerases are disclosed in the co-pending International PublicationNo. WO2014028429, the contents of which are incorporated herein byreference in their entirety.

In one aspect, the RNA polymerase which can be used in the synthesis ofthe polynucleotides of the present disclosure is a Syn5 RNA polymerase.(see Zhu et al. Nucleic Acids Research 2013, doi:10.1093/nar/gkt1193,which is herein incorporated by reference in its entirety). The Syn5 RNApolymerase was recently characterized from marine cyanophage Syn5 by Zhuet al. where they also identified the promoter sequence (see Zhu et al.Nucleic Acids Research 2013, the contents of which is hereinincorporated by reference in its entirety). Zhu et al. found that Syn5RNA polymerase catalyzed RNA synthesis over a wider range oftemperatures and salinity as compared to T7 RNA polymerase.Additionally, the requirement for the initiating nucleotide at thepromoter was found to be less stringent for Syn5 RNA polymerase ascompared to the T7 RNA polymerase making Syn5 RNA polymerase promisingfor RNA synthesis.

In one aspect, a Syn5 RNA polymerase can be used in the synthesis of thepolynucleotides described herein. As a non-limiting example, a Syn5 RNApolymerase can be used in the synthesis of the polynucleotide requiringa precise 3′-terminus.

In one aspect, a Syn5 promoter can be used in the synthesis of thepolynucleotides. As a non-limiting example, the Syn5 promoter can be5′-ATTGGGCACCCGTAAGGG-3′ (SEQ ID NO: 47) as described by Zhu et al.(Nucleic Acids Research 2013).

In one aspect, a Syn5 RNA polymerase can be used in the synthesis ofpolynucleotides comprising at least one chemical modification describedherein and/or known in the art (see e.g., the incorporation ofpseudo-UTP and 5Me-CTP described in Zhu et al. Nucleic Acids Research2013).

In one aspect, the polynucleotides described herein can be synthesizedusing a Syn5 RNA polymerase which has been purified using modified andimproved purification procedure described by Zhu et al. (Nucleic AcidsResearch 2013).

Various tools in genetic engineering are based on the enzymaticamplification of a target gene which acts as a template. For the studyof sequences of individual genes or specific regions of interest andother research needs, it is necessary to generate multiple copies of atarget gene from a small sample of polynucleotides or nucleic acids.Such methods can be applied in the manufacture of the polynucleotides ofthe disclosure.

For example, polymerase chain reaction (PCR), strand displacementamplification (SDA), nucleic acid sequence-based amplification (NASBA),also called transcription mediated amplification (TMA), and/orrolling-circle amplification (RCA) can be utilized in the manufacture ofone or more regions of the polynucleotides of the present invention.

Assembling polynucleotides or nucleic acids by a ligase is also widelyused.

b. Chemical Synthesis

Standard methods can be applied to synthesize an isolated polynucleotidesequence encoding an isolated polypeptide of interest, such as apolynucleotide of the disclosure. For example, a single DNA or RNAoligomer containing a codon-optimized nucleotide sequence coding for theparticular isolated polypeptide can be synthesized. In other aspects,several small oligonucleotides coding for portions of the desiredpolypeptide can be synthesized and then ligated. In some aspects, theindividual oligonucleotides typically contain 5′ or 3′ overhangs forcomplementary assembly.

A polynucleotide disclosed herein (e.g., a RNA, e.g., an mRNA) can bechemically synthesized using chemical synthesis methods and potentialnucleobase substitutions known in the art. See, for example,International Publication Nos. WO2014093924, WO2013052523; WO2013039857,WO2012135805, WO2013151671; U.S. Publ. No. US20130115272; or U.S. Pat.No. 8,999,380 or 8,710,200, all of which are herein incorporated byreference in their entireties.

c. Purification of Polynucleotides Encoding Tethered IL-12 Polypeptides

Purification of the polynucleotides described herein can include, but isnot limited to, polynucleotide clean-up, quality assurance and qualitycontrol. Clean-up can be performed by methods known in the arts such as,but not limited to, AGENCOURT® beads (Beckman Coulter Genomics, Danvers,Mass.), poly-T beads, LNA™ oligo-T capture probes (EXIQON® Inc.,Vedbaek, Denmark) or HPLC based purification methods such as, but notlimited to, strong anion exchange HPLC, weak anion exchange HPLC,reverse phase HPLC (RP-HPLC), and hydrophobic interaction HPLC(HIC-HPLC).

The term “purified” when used in relation to a polynucleotide such as a“purified polynucleotide” refers to one that is separated from at leastone contaminant. As used herein, a “contaminant” is any substance thatmakes another unfit, impure or inferior. Thus, a purified polynucleotide(e.g., DNA and RNA) is present in a form or setting different from thatin which it is found in nature, or a form or setting different from thatwhich existed prior to subjecting it to a treatment or purificationmethod.

In some embodiments, purification of a polynucleotide of the disclosureremoves impurities that can reduce or remove an unwanted immuneresponse, e.g., reducing cytokine activity.

In some embodiments, the polynucleotide of the disclosure is purifiedprior to administration using column chromatography (e.g., strong anionexchange HPLC, weak anion exchange HPLC, reverse phase HPLC (RP-HPLC),and hydrophobic interaction HPLC (HIC-HPLC), or (LCMS)).

In some embodiments, the polynucleotide of the disclosure purified usingcolumn chromatography (e.g., strong anion exchange HPLC, weak anionexchange HPLC, reverse phase HPLC (RP-HPLC, hydrophobic interaction HPLC(HIC-HPLC), or (LCMS)) presents increased expression of the encodedIL-12 protein compared to the expression level obtained with the samepolynucleotide of the present disclosure purified by a differentpurification method.

In some embodiments, a column chromatography (e.g., strong anionexchange HPLC, weak anion exchange HPLC, reverse phase HPLC (RP-HPLC),hydrophobic interaction HPLC (HIC-HPLC), or (LCMS)) purifiedpolynucleotide comprises a nucleotide sequence encoding an IL-12polypeptide comprising one or more of the point mutations known in theart.

In some embodiments, the use of RP-HPLC purified polynucleotideincreases IL-12 protein expression levels in cells when introduced intothose cells, e.g., by 10-100%, i.e., at least about 10%, at least about20%, at least about 25%, at least about 30%, at least about 35%, atleast about 40%, at least about 45%, at least about 50%, at least about55%, at least about 60%, at least about 65%, at least about 70%, atleast about 75%, at least about 80%, at least about 90%, at least about95%, or at least about 100% with respect to the expression levels ofIL-12 protein in the cells before the RP-HPLC purified polynucleotidewas introduced in the cells, or after a non-RP-HPLC purifiedpolynucleotide was introduced in the cells.

In some embodiments, the use of RP-HPLC purified polynucleotideincreases functional IL-12 protein expression levels in cells whenintroduced into those cells, e.g., by 10-100%, i.e., at least about 10%,at least about 20%, at least about 25%, at least about 30%, at leastabout 35%, at least about 40%, at least about 45%, at least about 50%,at least about 55%, at least about 60%, at least about 65%, at leastabout 70%, at least about 75%, at least about 80%, at least about 90%,at least about 95%, or at least about 100% with respect to thefunctional expression levels of IL-12 protein in the cells before theRP-HPLC purified polynucleotide was introduced in the cells, or after anon-RP-HPLC purified polynucleotide was introduced in the cells.

In some embodiments, the use of RP-HPLC purified polynucleotideincreases detectable IL-12 activity in cells when introduced into thosecells, e.g., by 10-100%, i.e., at least about 10%, at least about 20%,at least about 25%, at least about 30%, at least about 35%, at leastabout 40%, at least about 45%, at least about 50%, at least about 55%,at least about 60%, at least about 65%, at least about 70%, at leastabout 75%, at least about 80%, at least about 90%, at least about 95%,or at least about 100% with respect to the activity levels of functionalIL-12 in the cells before the RP-HPLC purified polynucleotide wasintroduced in the cells, or after a non-RP-HPLC purified polynucleotidewas introduced in the cells.

In some embodiments, the purified polynucleotide is at least about 80%pure, at least about 85% pure, at least about 90% pure, at least about95% pure, at least about 96% pure, at least about 97% pure, at leastabout 98% pure, at least about 99% pure, or about 100% pure.

A quality assurance and/or quality control check can be conducted usingmethods such as, but not limited to, gel electrophoresis, UV absorbance,or analytical HPLC. In another embodiment, the polynucleotide can besequenced by methods including, but not limited toreverse-transcriptase-PCR.

d. Quantification of Expressed Polynucleotides Encoding Tethered IL-12Polypeptides

In some embodiments, the polynucleotides of the present disclosure,their expression products, as well as degradation products andmetabolites can be quantified according to methods known in the art.

In some embodiments, the polynucleotides of the present disclosure canbe quantified in exosomes or when derived from one or more bodily fluid.As used herein “bodily fluids” include peripheral blood, serum, plasma,ascites, urine, cerebrospinal fluid (CSF), sputum, saliva, bone marrow,synovial fluid, aqueous humor, amniotic fluid, cerumen, breast milk,broncheoalveolar lavage fluid, semen, prostatic fluid, cowper's fluid orpre-ejaculatory fluid, sweat, fecal matter, hair, tears, cyst fluid,pleural and peritoneal fluid, pericardial fluid, lymph, chyme, chyle,bile, interstitial fluid, menses, pus, sebum, vomit, vaginal secretions,mucosal secretion, stool water, pancreatic juice, lavage fluids fromsinus cavities, bronchopulmonary aspirates, blastocyl cavity fluid, andumbilical cord blood. Alternatively, exosomes can be retrieved from anorgan selected from the group consisting of lung, heart, pancreas,stomach, intestine, bladder, kidney, ovary, testis, skin, colon, breast,prostate, brain, esophagus, liver, and placenta.

In the exosome quantification method, a sample of not more than 2 mL isobtained from the subject and the exosomes isolated by size exclusionchromatography, density gradient centrifugation, differentialcentrifugation, nanomembrane ultrafiltration, immunoabsorbent capture,affinity purification, microfluidic separation, or combinations thereof.In the analysis, the level or concentration of a polynucleotide can bean expression level, presence, absence, truncation or alteration of theadministered construct. It is advantageous to correlate the level withone or more clinical phenotypes or with an assay for a human diseasebiomarker.

The assay can be performed using construct specific probes, cytometry,qRT-PCR, real-time PCR, PCR, flow cytometry, electrophoresis, massspectrometry, or combinations thereof while the exosomes can be isolatedusing immunohistochemical methods such as enzyme linked immunosorbentassay (ELISA) methods. Exosomes may also be isolated by size exclusionchromatography, density gradient centrifugation, differentialcentrifugation, nanomembrane ultrafiltration, immunoabsorbent capture,affinity purification, microfluidic separation, or combinations thereof.

These methods afford the investigator the ability to monitor, in realtime, the level of polynucleotides remaining or delivered. This ispossible because the polynucleotides of the present disclosure differfrom the endogenous forms due to the structural or chemicalmodifications. In some embodiments, the polynucleotide can be quantifiedusing methods such as, but not limited to, ultraviolet visiblespectroscopy (UV/Vis). A non-limiting example of a UV/Vis spectrometeris a NANODROP® spectrometer (ThermoFisher, Waltham, Mass.). Thequantified polynucleotide can be analyzed in order to determine if thepolynucleotide can be of proper size, check that no degradation of thepolynucleotide has occurred. Degradation of the polynucleotide can bechecked by methods such as, but not limited to, agarose gelelectrophoresis, HPLC based purification methods such as, but notlimited to, strong anion exchange HPLC, weak anion exchange HPLC,reverse phase HPLC (RP-HPLC), and hydrophobic interaction HPLC(HIC-HPLC), liquid chromatography-mass spectrometry (LCMS), capillaryelectrophoresis (CE) and capillary gel electrophoresis (CGE).

24. Pharmaceutical Compositions and Formulations

The present disclosure provides pharmaceutical compositions andformulations that comprise any of the polynucleotides described above.In some embodiments, the composition or formulation further comprises adelivery agent.

In some embodiments, the composition or formulation can contain apolynucleotide comprising a sequence optimized nucleic acid sequencedisclosed herein which encodes a tethered IL-12 polypeptide comprisingan IL-12 polypeptide as disclosed herein and a membrane domain asdisclosed herein. In some embodiments, the composition or formulationcan contain a polynucleotide (e.g., a RNA, e.g., an mRNA) comprising anucleic acid sequence (e.g., an ORF) having significant sequenceidentity to a sequence optimized nucleic acid sequence disclosed hereinwhich encodes an IL-12 polypeptide and/or a sequence optimized nucleicacid sequence disclosed herein which encodes a membrane domain. In someembodiments, the polynucleotide further comprises a miRNA binding site,e.g., a miRNA binding site that binds miR-122.

Pharmaceutical compositions or formulation can optionally comprise oneor more additional active substances, e.g., therapeutically and/orprophylactically active substances. Pharmaceutical compositions orformulation of the present disclosure can be sterile and/orpyrogen-free. General considerations in the formulation and/ormanufacture of pharmaceutical agents can be found, for example, inRemington: The Science and Practice of Pharmacy 21^(st) ed., LippincottWilliams & Wilkins, 2005 (incorporated herein by reference in itsentirety). In some embodiments, compositions are administered to humans,human patients or subjects. For the purposes of the present disclosure,the phrase “active ingredient” generally refers to polynucleotides to bedelivered as described herein. Formulations and pharmaceuticalcompositions described herein can be prepared by any method known orhereafter developed in the art of pharmacology. In general, suchpreparatory methods include the step of associating the activeingredient with an excipient and/or one or more other accessoryingredients, and then, if necessary and/or desirable, dividing, shapingand/or packaging the product into a desired single- or multi-dose unit.

A pharmaceutical composition or formulation in accordance with thepresent disclosure can be prepared, packaged, and/or sold in bulk, as asingle unit dose, and/or as a plurality of single unit doses. As usedherein, a “unit dose” refers to a discrete amount of the pharmaceuticalcomposition comprising a predetermined amount of the active ingredient.The amount of the active ingredient is generally equal to the dosage ofthe active ingredient that would be administered to a subject and/or aconvenient fraction of such a dosage such as, for example, one-half orone-third of such a dosage. Relative amounts of the active ingredient,the pharmaceutically acceptable excipient, and/or any additionalingredients in a pharmaceutical composition in accordance with thepresent disclosure may vary, depending upon the identity, size, and/orcondition of the subject being treated and further depending upon theroute by which the composition is to be administered.

In some embodiments, the compositions and formulations described hereincan contain at least one polynucleotide of the disclosure. As anon-limiting example, the composition or formulation can contain 1, 2,3, 4 or 5 polynucleotides of the disclosure. In some embodiments, thecompositions or formulations described herein can comprise more than onetype of polynucleotide. In some embodiments, the composition orformulation can comprise a polynucleotide in linear and circular form.In another embodiment, the composition or formulation can comprise acircular polynucleotide and an IVT polynucleotide. In yet anotherembodiment, the composition or formulation can comprise an IVTpolynucleotide, a chimeric polynucleotide and a circular polynucleotide.

Although the descriptions of pharmaceutical compositions andformulations provided herein are principally directed to pharmaceuticalcompositions and formulations that are suitable for administration tohumans, it will be understood by the skilled artisan that suchcompositions are generally suitable for administration to any otheranimal, e.g., to non-human animals, e.g. non-human mammals.

The present disclosure provides pharmaceutical formulations thatcomprise a polynucleotide described herein. The polynucleotidesdescribed herein can be formulated using one or more excipients to:

(1) increase stability; (2) increase cell transfection; (3) permit thesustained or delayed release (e.g., from a depot formulation of thepolynucleotide); (4) alter the biodistribution (e.g., target thepolynucleotide to specific tissues or cell types); (5) increase thetranslation of encoded protein in vivo; and/or (6) alter the releaseprofile of encoded protein in vivo. In some embodiments, thepharmaceutical formulation further comprises a delivery agent, (e.g., acompound having the Formula (I)).

A pharmaceutically acceptable excipient, as used herein, includes, butare not limited to, any and all solvents, dispersion media, or otherliquid vehicles, dispersion or suspension aids, diluents, granulatingand/or dispersing agents, surface active agents, isotonic agents,thickening or emulsifying agents, preservatives, binders, lubricants oroil, coloring, sweetening or flavoring agents, stabilizers,antioxidants, antimicrobial or antifungal agents, osmolality adjustingagents, pH adjusting agents, buffers, chelants, cyoprotectants, and/orbulking agents, as suited to the particular dosage form desired. Variousexcipients for formulating pharmaceutical compositions and techniquesfor preparing the composition are known in the art (see Remington: TheScience and Practice of Pharmacy, 21st Edition, A. R. Gennaro(Lippincott, Williams & Wilkins, Baltimore, Md., 2006; incorporatedherein by reference in its entirety).

Exemplary diluents include, but are not limited to, calcium or sodiumcarbonate, calcium phosphate, calcium hydrogen phosphate, sodiumphosphate, lactose, sucrose, cellulose, microcrystalline cellulose,kaolin, mannitol, sorbitol, etc., and/or combinations thereof.

Exemplary granulating and/or dispersing agents include, but are notlimited to, starches, pregelatinized starches, or microcrystallinestarch, alginic acid, guar gum, agar, poly(vinyl-pyrrolidone),(providone), cross-linked poly(vinyl-pyrrolidone) (crospovidone),cellulose, methylcellulose, carboxymethyl cellulose, cross-linked sodiumcarboxymethyl cellulose (croscarmellose), magnesium aluminum silicate(VEEGUMg), sodium lauryl sulfate, etc., and/or combinations thereof.

Exemplary surface active agents and/or emulsifiers include, but are notlimited to, natural emulsifiers (e.g., acacia, agar, alginic acid,sodium alginate, tragacanth, chondrux, cholesterol, xanthan, pectin,gelatin, egg yolk, casein, wool fat, cholesterol, wax, and lecithin),sorbitan fatty acid esters (e.g., polyoxyethylene sorbitan monooleate[TWEEN®80], sorbitan monopalmitate [SPAN®40], glyceryl monooleate,polyoxyethylene esters, polyethylene glycol fatty acid esters (e.g.,CREMOPHOR®), polyoxyethylene ethers (e.g., polyoxyethylene lauryl ether[BRIPID30]), PLUORINC®F 68, POLOXAMER®188, etc. and/or combinationsthereof.

Exemplary binding agents include, but are not limited to, starch,gelatin, sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses,lactose, lactitol, mannitol), amino acids (e.g., glycine), natural andsynthetic gums (e.g., acacia, sodium alginate), ethylcellulose,hydroxyethylcellulose, hydroxypropyl methylcellulose, etc., andcombinations thereof.

Oxidation is a potential degradation pathway for mRNA, especially forliquid mRNA formulations. In order to prevent oxidation, antioxidantscan be added to the formulations. Exemplary antioxidants include, butare not limited to, alpha tocopherol, ascorbic acid, acorbyl palmitate,benzyl alcohol, butylated hydroxyanisole, m-cresol, methionine,butylated hydroxytoluene, monothioglycerol, sodium or potassiummetabisulfite, propionic acid, propyl gallate, sodium ascorbate, etc.,and combinations thereof.

Exemplary chelating agents include, but are not limited to,ethylenediaminetetraacetic acid (EDTA), citric acid monohydrate,disodium edetate, fumaric acid, malic acid, phosphoric acid, sodiumedetate, tartaric acid, trisodium edetate, etc., and combinationsthereof.

Exemplary antimicrobial or antifungal agents include, but are notlimited to, benzalkonium chloride, benzethonium chloride, methylparaben, ethyl paraben, propyl paraben, butyl paraben, benzoic acid,hydroxybenzoic acid, potassium or sodium benzoate, potassium or sodiumsorbate, sodium propionate, sorbic acid, etc., and combinations thereof.

Exemplary preservatives include, but are not limited to, vitamin A,vitamin C, vitamin E, beta-carotene, citric acid, ascorbic acid,butylated hydroxyanisol, ethylenediamine, sodium lauryl sulfate (SLS),sodium lauryl ether sulfate (SLES), etc., and combinations thereof

In some embodiments, the pH of polynucleotide solutions are maintainedbetween pH 5 and pH 8 to improve stability. Exemplary buffers to controlpH can include, but are not limited to sodium phosphate, sodium citrate,sodium succinate, histidine (or histidine-HCl), sodium malate, sodiumcarbonate, etc., and/or combinations thereof.

Exemplary lubricating agents include, but are not limited to, magnesiumstearate, calcium stearate, stearic acid, silica, talc, malt,hydrogenated vegetable oils, polyethylene glycol, sodium benzoate,sodium or magnesium lauryl sulfate, etc., and combinations thereof.

The pharmaceutical composition or formulation described here may containa cyroprotectant to stabilize a polynucleotide described herein duringfreezing. Exemplary cryoprotectants include, but are not limited tomannitol, sucrose, trehalose, lactose, glycerol, dextrose, etc., andcombinations thereof.

The pharmaceutical composition or formulation described here may containa bulking agent in lyophilized polynucleotide formulations to yield a“pharmaceutically elegant” cake, stabilize the lyophilizedpolynucleotides during long term (e.g., 36 month) storage. Exemplarybulking agents of the present disclosure can include, but are notlimited to sucrose, trehalose, mannitol, glycine, lactose, raffinose,and combinations thereof.

In some embodiments, the pharmaceutical composition or formulationfurther comprises a delivery agent. The delivery agent of the presentdisclosure can include, without limitation, liposomes, lipidnanoparticles, lipidoids, polymers, lipoplexes, microvesicles, exosomes,peptides, proteins, cells transfected with polynucleotides,hyaluronidase, nanoparticle mimics, nanotubes, conjugates, andcombinations thereof.

25. Delivery Agents

a. Lipid Compound

The present disclosure provides pharmaceutical compositions withadvantageous properties. In particular, the present application providespharmaceutical compositions comprising:

-   -   (a) a polynucleotide comprising a nucleotide sequence encoding a        tethered IL-12 polypeptide as disclosed herein; and    -   (b) a delivery agent.

In one embodiment, the delivery agent for the present disclosure is alipid nanoparticle. In another embodiment, the delivery agent comprisesformula (I):

In other embodiments, the delivery agent for the present disclosurecomprises any one or more compounds disclosed in InternationalApplication No. PCT/US2016/052352, filed on Sep. 16, 2016 and publishedas WO 2017/2017/049245, which is incorporated herein by reference in itsentirety.

Amine moieties of the lipid compounds disclosed herein can be protonatedunder certain conditions. For example, the central amine moiety of alipid according to formula (I) is typically protonated (i.e., positivelycharged) at a pH below the pKa of the amino moiety and is substantiallynot charged at a pH above the pKa. Such lipids may be referred toionizable amino lipids.

In one specific embodiment, the ionizable amino lipid is the compound offormula (I). In some embodiments, the amount the ionizable amino lipid,e.g., compound of formula (I), ranges from about 1 mol % to 99 mol % inthe lipid composition.

In one embodiment, the amount of the ionizable amino lipid, e.g.,compound of formula (I), is at least about 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45,46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63,64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81,82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or99 mol % in the lipid composition.

In one embodiment, the amount of the ionizable amino lipid, e.g., thecompound of formula (I), ranges from about 30 mol % to about 70 mol %,from about 35 mol % to about 65 mol %, from about 40 mol % to about 60mol %, and from about 45 mol % to about 55 mol % in the lipidcomposition.

In one specific embodiment, the amount of the compound of formula (I) isabout 50 mol % in the lipid composition.

In addition to the compound of formula (I), the lipid composition of thepharmaceutical compositions disclosed herein can comprise additionalcomponents such as phospholipids, structural lipids, quaternary aminecompounds, PEG-lipids, and any combination thereof.

b. Additional Components in the Lipid Composition

(i) Phospholipids

The lipid composition of the pharmaceutical composition disclosed hereincan comprise one or more phospholipids, for example, one or moresaturated or (poly)unsaturated phospholipids or a combination thereof.In general, phospholipids comprise a phospholipid moiety and one or morefatty acid moieties.

A phospholipid moiety may be selected, for example, from thenon-limiting group consisting of phosphatidyl choline, phosphatidylethanolamine, phosphatidyl glycerol, phosphatidyl serine, phosphatidicacid, 2-lysophosphatidyl choline, and a sphingomyelin.

A fatty acid moiety may be selected, for example, from the non-limitinggroup consisting of lauric acid, myristic acid, myristoleic acid,palmitic acid, palmitoleic acid, stearic acid, oleic acid, linoleicacid, alpha-linolenic acid, erucic acid, phytanoic acid, arachidic acid,arachidonic acid, eicosapentaenoic acid, behenic acid, docosapentaenoicacid, and docosahexaenoic acid.

Particular phospholipids may facilitate fusion to a membrane. Forexample, a cationic phospholipid may interact with one or morenegatively charged phospholipids of a membrane (e.g., a cellular orintracellular membrane). Fusion of a phospholipid to a membrane mayallow one or more elements (e.g., a therapeutic agent) of alipid-containing composition (e.g., LNPs) to pass through the membranepermitting, e.g., delivery of the one or more elements to a targettissue (e.g., tumoral tissue).

Non-natural phospholipid species including natural species withmodifications and substitutions including branching, oxidation,cyclization, and alkynes are also contemplated. For example, aphospholipid may be functionalized with or cross-linked to one or morealkynes (e.g., an alkenyl group in which one or more double bonds isreplaced with a triple bond). Under appropriate reaction conditions, analkyne group may undergo a copper-catalyzed cycloaddition upon exposureto an azide. Such reactions may be useful in functionalizing a lipidbilayer of a nanoparticle composition to facilitate membrane permeationor cellular recognition or in conjugating a nanoparticle composition toa useful component such as a targeting or imaging moiety (e.g., a dye).

Phospholipids include, but are not limited to, glycerophospholipids suchas phosphatidylcholines, phosphatidylethanolamines, phosphatidylserines,phosphatidylinositols, phosphatidy glycerols, and phosphatidic acids.Phospholipids also include phosphosphingolipid, such as sphingomyelin.In some embodiments, a pharmaceutical composition for intratumoraldelivery disclosed herein can comprise more than one phospholipid. Whenmore than one phospholipid is used, such phospholipids can belong to thesame phospholipid class (e.g., MSPC and DSPC) or different classes(e.g., MSPC and MSPE).

Phospholipids may be of a symmetric or an asymmetric type. As usedherein, the term “symmetric phospholipid” includes glycerophospholipidshaving matching fatty acid moieties and sphingolipids in which thevariable fatty acid moiety and the hydrocarbon chain of the sphingosinebackbone include a comparable number of carbon atoms. As used herein,the term “asymmetric phospholipid” includes lysolipids,glycerophospholipids having different fatty acid moieties (e.g., fattyacid moieties with different numbers of carbon atoms and/orunsaturations (e.g., double bonds)), and sphingolipids in which thevariable fatty acid moiety and the hydrocarbon chain of the sphingosinebackbone include a dissimilar number of carbon atoms (e.g., the variablefatty acid moiety include at least two more carbon atoms than thehydrocarbon chain or at least two fewer carbon atoms than thehydrocarbon chain).

In some embodiments, the lipid composition of a pharmaceuticalcomposition disclosed herein comprises at least one symmetricphospholipid.

In some embodiments, the lipid composition of a pharmaceuticalcomposition disclosed herein comprises at least one symmetricphospholipid selected from the non-limiting group consisting of DLPC,DMPC, DOPC, DPPC, DSPC, DUPC, 18:0 Diether PC, DLnPC, DAPC, DHAPC, DOPE,4ME 16:0 PE, DSPE, DLPE, DLnPE, DAPE, DHAPE, DOPG, and any combinationthereof.

In some embodiment, the lipid composition of a pharmaceuticalcomposition disclosed herein comprises at least one asymmetricphospholipid selected from the group consisting of MPPC, MSPC, PMPC,PSPC, SMPC, SPPC, and any combination thereof. In some embodiments, theasymmetric phospholipid is1-myristoyl-2-stearoyl-sn-glycero-3-phosphocholine (MSPC). In aparticular embodiment, the asymmetric phospholipid is one or morephospholipid disclosed in International Application No. PCT/US17/27492,filed on Apr. 13, 2017, which is incorporated herein by reference in itsentireties.

In some embodiments, the lipid compositions disclosed herein may containone or more symmetric phospholipids, one or more asymmetricphospholipids, or a combination thereof. When multiple phospholipids arepresent, they can be present in equimolar ratios, or non-equimolarratios.

In one embodiment, the lipid composition of a pharmaceutical compositiondisclosed herein comprises a total amount of phospholipid (e.g., MSPC)which ranges from about 1 mol % to about 20 mol %, from about 5 mol % toabout 20 mol %, from about 10 mol % to about 20 mol %, from about 15 mol% to about 20 mol %, from about 1 mol % to about 15 mol %, from about 5mol % to about 15 mol %, from about 10 mol % to about 15 mol %, fromabout 5 mol % to about 10 mol % in the lipid composition. In oneembodiment, the amount of the phospholipid is from about 8 mol % toabout 15 mol % in the lipid composition. In one embodiment, the amountof the phospholipid (e.g., MSPC) is about 10 mol % in the lipidcomposition.

In some aspects, the amount of a specific phospholipid (e.g., MSPC) isat least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,17, 18, 19 or 20 mol % in the lipid composition.

(ii) Quaternary Amine Compounds

The lipid composition of a pharmaceutical composition disclosed hereincan comprise one or more quaternary amine compounds (e.g., DOTAP). Theterm “quaternary amine compound” is used to include those compoundshaving one or more quaternary amine groups (e.g., trialkylamino groups)and permanently carrying a positive charge and existing in a form of asalt. For example, the one or more quaternary amine groups can bepresent in a lipid or a polymer (e.g., PEG). In some embodiments, thequaternary amine compound comprises (1) a quaternary amine group and (2)at least one hydrophobic tail group comprising (i) a hydrocarbon chain,linear or branched, and saturated or unsaturated, and (ii) optionally anether, ester, carbonyl, or ketal linkage between the quaternary aminegroup and the hydrocarbon chain. In some embodiments, the quaternaryamine group can be a trimethylammonium group. In some embodiments, thequaternary amine compound comprises two identical hydrocarbon chains. Insome embodiments, the quaternary amine compound comprises two differenthydrocarbon chains.

In some embodiments, the lipid composition of a pharmaceuticalcomposition disclosed herein comprises at least one quaternary aminecompound. In one embodiment, the quaternary amine compound is1,2-dioleoyl-3-trimethylammonium-propane (DOTAP). Quaternary aminecompounds are known in the art, such as those described in US2013/0245107 A1, US 2014/0363493 A1, U.S. Pat. No. 8,158,601, WO2015/123264 A1, and WO 2015/148247 A1, which are incorporated herein byreference in their entirety.

In one embodiment, the amount of the quaternary amine compound (e.g.,DOTAP) in the lipid composition disclosed herein ranges from about 0.01mol % to about 20 mol %.

In one embodiment, the amount of the quaternary amine compound (e.g.,DOTAP) in the lipid composition disclosed herein ranges from about 0.5mol % to about 20 mol %, from about 0.5 mol % to about 15 mol %, fromabout 0.5 mol % to about 10 mol %, from about 1 mol % to about 20 mol %,from about 1 mol % to about 15 mol %, from about 1 mol % to about 10 mol%, from about 2 mol % to about 20 mol %, from about 2 mol % to about 15mol %, from about 2 mol % to about 10 mol %, from about 3 mol % to about20 mol %, from about 3 mol % to about 15 mol %, from about 3 mol % toabout 10 mol %, from about 4 mol % to about 20 mol %, from about 4 mol %to about 15 mol %, from about 4 mol % to about 10 mol %, from about 5mol % to about 20 mol %, from about 5 mol % to about 15 mol %, fromabout 5 mol % to about 10 mol %, from about 6 mol % to about 20 mol %,from about 6 mol % to about 15 mol %, from about 6 mol % to about 10 mol%, from about 7 mol % to about 20 mol %, from about 7 mol % to about 15mol %, from about 7 mol % to about 10 mol %, from about 8 mol % to about20 mol %, from about 8 mol % to about 15 mol %, from about 8 mol % toabout 10 mol %, from about 9 mol % to about 20 mol %, from about 9 mol %to about 15 mol %, from about 9 mol % to about 10 mol %.

In one embodiment, the amount of the quaternary amine compound (e.g.,DOTAP) in the lipid composition disclosed herein ranges from about 5 mol% to about 10 mol %.

In one embodiment, the amount of the quaternary amine compound (e.g.,DOTAP) in the lipid composition disclosed herein is about 5 mol %. Inone embodiment, the amount of the quaternary amine compound (e.g.,DOTAP) in the lipid composition disclosed herein is about 10 mol %.

In some embodiments, the amount of the quaternary amine compound (e.g.,DOTAP) is at least about 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08,0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.5, 2, 2.5, 3,3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5,12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16, 16.5, 17, 17.5, 18, 18.5,19, 19.5 or 20 mol % in the lipid composition disclosed herein.

In some embodiments, the lipid composition of the pharmaceuticalcompositions disclosed herein comprises a compound of formula (I). Inone embodiment, the mole ratio of the compound of formula (I) to thequaternary amine compound (e.g., DOTA) is about 100:1 to about 2.5:1. Inone embodiment, the mole ratio of the compound of formula (I) to thequaternary amine compound (e.g., DOTAP) is about 90:1, about 80:1, about70:1, about 60:1, about 50:1, about 40:1, about 30:1, about 20:1, about15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1,or about 2.5:1. In one embodiment, the mole ratio of the compound offormula (I) to the quaternary amine compound (e.g., DOTAP) in the lipidcomposition disclosed herein is about 10:1.

In some aspects, the lipid composition of the pharmaceuticalcompositions disclosed herein does not comprise a quaternary aminecompound. In some aspects, the lipid composition of the pharmaceuticalcompositions disclosed herein does not comprise DOTAP.

(iii) Structural Lipids

The lipid composition of a pharmaceutical composition disclosed hereincan comprise one or more structural lipids. As used herein, the term“structural lipid” refers to sterols and also to lipids containingsterol moieties. In some embodiments, the structural lipid is selectedfrom the group consisting of cholesterol, fecosterol, sitosterol,ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine,tomatine, ursolic acid, alpha-tocopherol, and mixtures thereof. In someembodiments, the structural lipid is cholesterol.

In one embodiment, the amount of the structural lipid (e.g., an sterolsuch as cholesterol) in the lipid composition of a pharmaceuticalcomposition disclosed herein ranges from about 20 mol % to about 60 mol%, from about 25 mol % to about 55 mol %, from about 30 mol % to about50 mol %, or from about 35 mol % to about 45 mol %.

In one embodiment, the amount of the structural lipid (e.g., an sterolsuch as cholesterol) in the lipid composition disclosed herein rangesfrom about 25 mol % to about 30 mol %, from about 30 mol % to about 35mol %, or from about 35 mol % to about 40 mol %.

In one embodiment, the amount of the structural lipid (e.g., a sterolsuch as cholesterol) in the lipid composition disclosed herein is about23.5 mol %, about 28.5 mol %, about 33.5 mol %, or about 38.5 mol %.

In some embodiments, the amount of the structural lipid (e.g., an sterolsuch as cholesterol) in the lipid composition disclosed herein is atleast about 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34,35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52,53, 54, 55, 56, 57, 58, 59, or 60 mol %.

In some aspects, the lipid composition component of the pharmaceuticalcompositions for intratumoral delivery disclosed does not comprisecholesterol.

(iv) Polyethylene Glycol (PEG)-Lipids

The lipid composition of a pharmaceutical composition disclosed hereincan comprise one or more a polyethylene glycol (PEG) lipid.

As used herein, the term “PEG-lipid” refers to polyethylene glycol(PEG)-modified lipids. Non-limiting examples of PEG-lipids includePEG-modified phosphatidylethanolamine and phosphatidic acid,PEG-ceramide conjugates (e.g., PEG-CerC14 or PEG-CerC20), PEG-modifieddialkylamines and PEG-modified 1,2-diacyloxypropan-3-amines. Such lipidsare also referred to as PEGylated lipids. For example, a PEG lipid maybe PEG-c-DOMG, PEG-DMG, PEG-DLPE, PEG-DMPE, PEG-DPPC, or a PEG-DSPElipid.

In some embodiments, the PEG-lipid includes, but not limited to1,2-dimyristoyl-sn-glycerol methoxypolyethylene glycol (PEG-DMG),1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethyleneglycol)] (PEG-DSPE), PEG-disteryl glycerol (PEG-DSG), PEG-dipalmetoleyl,PEG-dioleyl, PEG-distearyl, PEG-diacylglycamide (PEG-DAG),PEG-dipalmitoyl phosphatidylethanolamine (PEG-DPPE), orPEG-1,2-dimyristyloxlpropyl-3-amine (PEG-c-DMA).

In one embodiment, the PEG-lipid is selected from the group consistingof a PEG-modified phosphatidylethanolamine, a PEG-modified phosphatidicacid, a PEG-modified ceramide, a PEG-modified dialkylamine, aPEG-modified diacylglycerol, a PEG-modified dialkylglycerol, andmixtures thereof.

In some embodiments, the lipid moiety of the PEG-lipids includes thosehaving lengths of from about C₁₄ to about C₂₂, preferably from about C₁₄to about C₁₆. In some embodiments, a PEG moiety, for example anmPEG-NH₂, has a size of about 1000, 2000, 5000, 10,000, 15,000 or 20,000daltons. In one embodiment, the PEG-lipid is PEG_(2k)-DMG.

In one embodiment, the lipid nanoparticles described herein may comprisea PEG lipid which is a non-diffusible PEG. Non-limiting examples ofnon-diffusible PEGs include PEG-DSG and PEG-DSPE.

PEG-lipids are known in the art, such as those described in U.S. Pat.No. 8,158,601 and International Publ. No. WO 2015/130584 A2, which areincorporated herein by reference in their entirety.

In one embodiment, the amount of PEG-lipid in the lipid composition of apharmaceutical composition disclosed herein ranges from about 0.1 mol %to about 5 mol %, from about 0.5 mol % to about 5 mol %, from about 1mol % to about 5 mol %, from about 1.5 mol % to about 5 mol %, fromabout 2 mol % to about 5 mol % mol %, from about 0.1 mol % to about 4mol %, from about 0.5 mol % to about 4 mol %, from about 1 mol % toabout 4 mol %, from about 1.5 mol % to about 4 mol %, from about 2 mol %to about 4 mol %, from about 0.1 mol % to about 3 mol %, from about 0.5mol % to about 3 mol %, from about 1 mol % to about 3 mol %, from about1.5 mol % to about 3 mol %, from about 2 mol % to about 3 mol %, fromabout 0.1 mol % to about 2 mol %, from about 0.5 mol % to about 2 mol %,from about 1 mol % to about 2 mol %, from about 1.5 mol % to about 2 mol%, from about 0.1 mol % to about 1.5 mol %, from about 0.5 mol % toabout 1.5 mol %, or from about 1 mol % to about 1.5 mol %.

In one embodiment, the amount of PEG-lipid in the lipid compositiondisclosed herein is about 1.5 mol %.

In one embodiment, the amount of PEG-lipid in the lipid compositiondisclosed herein is at least about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7,0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2,2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7,3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or 5 mol %.

In some aspects, the lipid composition of the pharmaceuticalcompositions disclosed herein does not comprise a PEG-lipid.

In some embodiments, the lipid composition disclosed herein comprises anionizable amino lipid, e.g., compound of formula (I), and an asymmetricphospholipid. In some embodiments, the lipid composition comprisescompound 18 and MSPC.

In some embodiments, the lipid composition disclosed herein comprises anionizable amino lipid, e.g., compound of formula (I), and a quaternaryamine compound. In some embodiments, the lipid composition comprisescompound 18 and DOTAP.

In some embodiments, the lipid composition disclosed herein comprises anionizable amino lipid, e.g., compound of formula (I), an asymmetricphospholipid, and a quaternary amine compound. In some embodiments, thelipid composition comprises compound 18, MSPC and DOTAP.

In one embodiment, the lipid composition comprises about 50 mol % of acompound of formula (I), about 10 mol % of DSPC or MSPC, about 33.5 mol% of cholesterol, about 1.5 mol % of PEG-DMG, and about 5 mol % ofDOTAP. In one embodiment, the lipid composition comprises about 50 mol %of a compound of formula (I), about 10 mol % of DSPC or MSPC, about 28.5mol % of cholesterol, about 1.5 mol % of PEG-DMG, and about 10 mol % ofDOTAP.

The components of the lipid nanoparticle may be tailored for optimaldelivery of the polynucleotides based on the desired outcome. As anon-limiting example, the lipid nanoparticle may comprise 40-60 mol % anionizable amino lipid (e.g., a compound of formula (I), 8-16 mol %phospholipid, 30-45 mol % cholesterol, 1-5 mol % PEG lipid, andoptionally 1-15 mol % quaternary amine compound.

In some embodiments, the lipid nanoparticle may comprise 45-65 mol % ofan ionizable amino lipid (e.g., a compound of formula (I)), 5-10 mol %phospholipid, 25-40 mol % cholesterol, 0.5-5 mol % PEG lipid, andoptionally 1-15 mol % quaternary amine compound.

Non-limiting examples of nucleic acid lipid particles are disclosed inU.S. Patent Publication No. 20140121263, herein incorporated byreference in its entirety.

(v) Other Ionizable Amino Lipids

The lipid composition of the pharmaceutical composition disclosed hereincan comprise one or more ionizable amino lipids in addition to a lipidaccording to formula (I).

Ionizable lipids may be selected from the non-limiting group consistingof

-   3-(didodecylamino)-N1,N1,4-tridodecyl-1-piperazineethanamine (KL10),-   N1-[2-(didodecylamino)ethyl]-N1,N4,N4-tridodecyl-1,4-piperazinediethanamine    (KL22),-   14,25-ditridecyl-15,18,21,24-tetraaza-octatriacontane (KL25),-   1,2-dilinoleyloxy-N,N-dimethylaminopropane (DLin-DMA),-   2,2-dilinoleyl-4-dimethylaminomethyl-[1,3]-dioxolane (DLin-K-DMA),-   heptatriaconta-6,9,28,31-tetraen-19-yl 4-(dimethylamino)butanoate    (DLin-MC3-DMA),-   2,2-dilinoleyl-4-(2-dimethylaminoethyl)[1,3]-dioxolane    (DLin-KC2-DMA),-   1,2-dioleyloxy-N,N-dimethylaminopropane (DODMA),    (13Z,165Z)-N,N-dimethyl-3-nonydocosa-13-16-dien-1-amine (L608),-   2-({8-[(3β)-cholest-5-en-3-yloxy]    octyl}oxy)-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]propan-1-amine    (Octyl-CLinDMA),-   (2R)-2-({8-[(3β)-cholest-5-en-3-yloxy]octyl}oxy)-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien    yloxy]propan-1-amine (Octyl-CLinDMA (2R)), and-   (2S)-2-({8-[(3β)-cholest-5-en-3-yloxy]    octyl}oxy)-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien    yloxy]propan-1-amine (Octyl-CLinDMA (2S)). In addition to these, an    ionizable amino lipid may also be a lipid including a cyclic amine    group.

Ionizable lipids can also be the compounds disclosed in InternationalPublication No. WO 2015/199952 A1, hereby incorporated by reference inits entirety.

(vi) Other Lipid Composition Components

The lipid composition of a pharmaceutical composition disclosed hereinmay include one or more components in addition to those described above.For example, the lipid composition may include one or more permeabilityenhancer molecules, carbohydrates, polymers, surface altering agents(e.g., surfactants), or other components. For example, a permeabilityenhancer molecule may be a molecule described by U.S. Patent ApplicationPublication No. 2005/0222064. Carbohydrates may include simple sugars(e.g., glucose) and polysaccharides (e.g., glycogen and derivatives andanalogs thereof). The lipid composition may include a buffer such as,but not limited to, citrate or phosphate at a pH of 7, salt and/orsugar. Salt and/or sugar may be included in the formulations describedherein for isotonicity.

A polymer may be included in and/or used to encapsulate or partiallyencapsulate a pharmaceutical composition disclosed herein (e.g., apharmaceutical composition in lipid nanoparticle form). A polymer may bebiodegradable and/or biocompatible. A polymer may be selected from, butis not limited to, polyamines, polyethers, polyamides, polyesters,polycarbamates, polyureas, polycarbonates, polystyrenes, polyimides,polysulfones, polyurethanes, polyacetylenes, polyethylenes,polyethyleneimines, polyisocyanates, polyacrylates, polymethacrylates,polyacrylonitriles, and polyarylates.

The ratio between the lipid composition and the polynucleotide range canbe from about 10:1 to about 60:1 (wt/wt).

In some embodiments, the ratio between the lipid composition and thepolynucleotide can be about 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1,17:1, 18:1, 19:1, 20:1, 21:1, 22:1, 23:1, 24:1, 25:1, 26:1, 27:1, 28:1,29:1, 30:1, 31:1, 32:1, 33:1, 34:1, 35:1, 36:1, 37:1, 38:1, 39:1, 40:1,41:1, 42:1, 43:1, 44:1, 45:1, 46:1, 47:1, 48:1, 49:1, 50:1, 51:1, 52:1,53:1, 54:1, 55:1, 56:1, 57:1, 58:1, 59:1 or 60:1 (wt/wt). In someembodiments, the wt/wt ratio of the lipid composition to thepolynucleotide encoding a therapeutic agent is about 20:1 or about 15:1.

In some embodiments, the pharmaceutical composition disclosed herein cancontain more than one polypeptides. For example, a pharmaceuticalcomposition disclosed herein can contain two or more polynucleotides(e.g., RNA, e.g., mRNA).

In one embodiment, the lipid nanoparticles described herein may comprisepolynucleotides (e.g., mRNA) in a lipid:polynucleotide weight ratio of5:1, 10:1, 15:1, 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, 50:1, 55:1, 60:1 or70:1, or a range or any of these ratios such as, but not limited to, 5:1to about 10:1, from about 5:1 to about 15:1, from about 5:1 to about20:1, from about 5:1 to about 25:1, from about 5:1 to about 30:1, fromabout 5:1 to about 35:1, from about 5:1 to about 40:1, from about 5:1 toabout 45:1, from about 5:1 to about 50:1, from about 5:1 to about 55:1,from about 5:1 to about 60:1, from about 5:1 to about 70:1, from about10:1 to about 15:1, from about 10:1 to about 20:1, from about 10:1 toabout 25:1, from about 10:1 to about 30:1, from about 10:1 to about35:1, from about 10:1 to about 40:1, from about 10:1 to about 45:1, fromabout 10:1 to about 50:1, from about 10:1 to about 55:1, from about 10:1to about 60:1, from about 10:1 to about 70:1, from about 15:1 to about20:1, from about 15:1 to about 25:1, from about 15:1 to about 30:1, fromabout 15:1 to about 35:1, from about 15:1 to about 40:1, from about 15:1to about 45:1, from about 15:1 to about 50:1, from about 15:1 to about55:1, from about 15:1 to about 60:1 or from about 15:1 to about 70:1. Inone embodiment, the lipid nanoparticles described herein may comprisethe polynucleotide in a concentration from approximately 0.1 mg/ml to 2mg/ml such as, but not limited to, 0.1 mg/ml, 0.2 mg/ml, 0.3 mg/ml, 0.4mg/ml, 0.5 mg/ml, 0.6 mg/ml, 0.7 mg/ml, 0.8 mg/ml, 0.9 mg/ml, 1.0 mg/ml,1.1 mg/ml, 1.2 mg/ml, 1.3 mg/ml, 1.4 mg/ml, 1.5 mg/ml, 1.6 mg/ml, 1.7mg/ml, 1.8 mg/ml, 1.9 mg/ml, 2.0 mg/ml or greater than 2.0 mg/ml.

In one embodiment, formulations comprising the polynucleotides and lipidnanoparticles described herein may comprise 0.15 mg/ml to 2 mg/ml of thepolynucleotide described herein (e.g., mRNA). In some embodiments, theformulation may further comprise 10 mM of citrate buffer and theformulation may additionally comprise up to 10% w/w of sucrose (e.g., atleast 1% w/w, at least 2% w/w/, at least 3% w/w, at least 4% w/w, atleast 5% w/w, at least 6% w/w, at least 7% w/w, at least 8% w/w, atleast 9% w/w or 10% w/w).

(vii) Nanoparticle Compositions

In some embodiments, the pharmaceutical compositions disclosed hereinare formulated as lipid nanoparticles (LNP). Accordingly, the presentdisclosure also provides nanoparticle compositions comprising (i) alipid composition comprising a delivery agent such as a compound offormula (I) as described herein, and (ii) a polynucleotide of thedisclosure. In such nanoparticle composition, the lipid compositiondisclosed herein can encapsulate the polynucleotide of the disclosure.

Nanoparticle compositions are typically sized on the order ofmicrometers or smaller and may include a lipid bilayer. Nanoparticlecompositions encompass lipid nanoparticles (LNPs), liposomes (e.g.,lipid vesicles), and lipoplexes. For example, a nanoparticle compositionmay be a liposome having a lipid bilayer with a diameter of 500 nm orless.

Nanoparticle compositions include, for example, lipid nanoparticles(LNPs), liposomes, and lipoplexes. In some embodiments, nanoparticlecompositions are vesicles including one or more lipid bilayers. Incertain embodiments, a nanoparticle composition includes two or moreconcentric bilayers separated by aqueous compartments. Lipid bilayersmay be functionalized and/or crosslinked to one another. Lipid bilayersmay include one or more ligands, proteins, or channels.

Nanoparticle compositions of the present disclosure comprise at leastone compound according to formula (I). Nanoparticle compositions canalso include a variety of other components. For example, thenanoparticle composition may include one or more other lipids inaddition to a lipid according to formula (I), such as (i) at least onephospholipid, (ii) at least one quaternary amine compound, (iii) atleast one structural lipid, (iv) at least one PEG-lipid, or (v) anycombination thereof.

In some embodiments, the nanoparticle composition comprises a compoundof formula (I) and a phospholipid (e.g., DSPC or MSPC). In someembodiments, the nanoparticle composition comprises a compound offormula (I), a phospholipid (e.g., DSPC or MSPC), and a quaternary aminecompound (e.g., DOTAP). In some embodiments, the nanoparticlecomposition comprises a compound of formula (I), and a quaternary aminecompound (e.g., DOTAP).

In one embodiment, the nanoparticle composition comprises (1) a lipidcomposition comprising about 50 mole % of a compound of formula (I);about 10 mole % of DSPC or MSPC; about 33.5 mole % of cholesterol; about1.5 mole % of PEG-DMG (e.g., PEG2k-DMG); about 5 mole % of DOTAP; and(2) a polynucleotide.

In one embodiment, the nanoparticle composition comprises (1) a lipidcomposition comprising about 50 mole % of a compound of formula (I);about 10 mole % of DSPC or MSPC; about 28.5 mole % of cholesterol; about1.5 mole % of PEG-DMG (e.g., PEG2k-DMG); about 10 mole % of DOTAP; and(2) a polynucleotide.

In one embodiment, the nanoparticle composition comprises (1) a lipidcomposition comprising about 50 mole % of a compound of formula (I);about 10 mole % of DSPC or MSPC; about 23.5 mole % of cholesterol; about1.5 mole % of PEG-DMG (e.g., PEG2k-DMG); about 15 mole % of DOTAP; and(2) a polynucleotide.

Nanoparticle compositions may be characterized by a variety of methods.For example, microscopy (e.g., transmission electron microscopy orscanning electron microscopy) may be used to examine the morphology andsize distribution of a nanoparticle composition. Dynamic lightscattering or potentiometry (e.g., potentiometric titrations) may beused to measure zeta potentials. Dynamic light scattering may also beutilized to determine particle sizes. Instruments such as the ZetasizerNano ZS (Malvern Instruments Ltd, Malvern, Worcestershire, UK) may alsobe used to measure multiple characteristics of a nanoparticlecomposition, such as particle size, polydispersity index, and zetapotential.

The size of the nanoparticles can help counter biological reactions suchas, but not limited to, inflammation, or can increase the biologicaleffect of the polynucleotide.

As used herein, “size” or “mean size” in the context of nanoparticlecompositions refers to the mean diameter of a nanoparticle composition.

In one embodiment, the polynucleotides of the disclosure are formulatedin lipid nanoparticles having a diameter from about 10 to about 100 nmsuch as, but not limited to, about 10 to about 20 nm, about 10 to about30 nm, about 10 to about 40 nm, about 10 to about 50 nm, about 10 toabout 60 nm, about 10 to about 70 nm, about 10 to about 80 nm, about 10to about 90 nm, about 20 to about 30 nm, about 20 to about 40 nm, about20 to about 50 nm, about 20 to about 60 nm, about 20 to about 70 nm,about 20 to about 80 nm, about 20 to about 90 nm, about 20 to about 100nm, about 30 to about 40 nm, about 30 to about 50 nm, about 30 to about60 nm, about 30 to about 70 nm, about 30 to about 80 nm, about 30 toabout 90 nm, about 30 to about 100 nm, about 40 to about 50 nm, about 40to about 60 nm, about 40 to about 70 nm, about 40 to about 80 nm, about40 to about 90 nm, about 40 to about 100 nm, about 50 to about 60 nm,about 50 to about 70 nm, about 50 to about 80 nm, about 50 to about 90nm, about 50 to about 100 nm, about 60 to about 70 nm, about 60 to about80 nm, about 60 to about 90 nm, about 60 to about 100 nm, about 70 toabout 80 nm, about 70 to about 90 nm, about 70 to about 100 nm, about 80to about 90 nm, about 80 to about 100 nm and/or about 90 to about 100nm.

In one embodiment, the nanoparticles have a diameter from about 10 to500 nm. In one embodiment, the nanoparticle has a diameter greater than100 nm, greater than 150 nm, greater than 200 nm, greater than 250 nm,greater than 300 nm, greater than 350 nm, greater than 400 nm, greaterthan 450 nm, greater than 500 nm, greater than 550 nm, greater than 600nm, greater than 650 nm, greater than 700 nm, greater than 750 nm,greater than 800 nm, greater than 850 nm, greater than 900 nm, greaterthan 950 nm or greater than 1000 nm.

In some embodiments, the largest dimension of a nanoparticle compositionis 1 μm or shorter (e.g., 1 μm, 900 nm, 800 nm, 700 nm, 600 nm, 500 nm,400 nm, 300 nm, 200 nm, 175 nm, 150 nm, 125 nm, 100 nm, 75 nm, 50 nm, orshorter).

A nanoparticle composition may be relatively homogenous. Apolydispersity index may be used to indicate the homogeneity of ananoparticle composition, e.g., the particle size distribution of thenanoparticle composition. A small (e.g., less than 0.3) polydispersityindex generally indicates a narrow particle size distribution. Ananoparticle composition may have a polydispersity index from about 0 toabout 0.25, such as 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08,0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20,0.21, 0.22, 0.23, 0.24, or 0.25. In some embodiments, the polydispersityindex of a nanoparticle composition disclosed herein may be from about0.10 to about 0.20.

The zeta potential of a nanoparticle composition may be used to indicatethe electrokinetic potential of the composition. For example, the zetapotential may describe the surface charge of a nanoparticle composition.Nanoparticle compositions with relatively low charges, positive ornegative, are generally desirable, as more highly charged species mayinteract undesirably with cells, tissues, and other elements in thebody. In some embodiments, the zeta potential of a nanoparticlecomposition disclosed herein may be from about −10 mV to about +20 mV,from about −10 mV to about +15 mV, from about 10 mV to about +10 mV,from about −10 mV to about +5 mV, from about −10 mV to about 0 mV, fromabout −10 mV to about −5 mV, from about −5 mV to about +20 mV, fromabout −5 mV to about +15 mV, from about −5 mV to about +10 mV, fromabout −5 mV to about +5 mV, from about −5 mV to about 0 mV, from about 0mV to about +20 mV, from about 0 mV to about +15 mV, from about 0 mV toabout +10 mV, from about 0 mV to about +5 mV, from about +5 mV to about+20 mV, from about +5 mV to about +15 mV, or from about +5 mV to about+10 mV.

In some embodiments, the zeta potential of the lipid nanoparticles maybe from about 0 mV to about 100 mV, from about 0 mV to about 90 mV, fromabout 0 mV to about 80 mV, from about 0 mV to about 70 mV, from about 0mV to about 60 mV, from about 0 mV to about 50 mV, from about 0 mV toabout 40 mV, from about 0 mV to about 30 mV, from about 0 mV to about 20mV, from about 0 mV to about 10 mV, from about 10 mV to about 100 mV,from about 10 mV to about 90 mV, from about 10 mV to about 80 mV, fromabout 10 mV to about 70 mV, from about 10 mV to about 60 mV, from about10 mV to about 50 mV, from about 10 mV to about 40 mV, from about 10 mVto about 30 mV, from about 10 mV to about 20 mV, from about 20 mV toabout 100 mV, from about 20 mV to about 90 mV, from about 20 mV to about80 mV, from about 20 mV to about 70 mV, from about 20 mV to about 60 mV,from about 20 mV to about 50 mV, from about 20 mV to about 40 mV, fromabout 20 mV to about 30 mV, from about 30 mV to about 100 mV, from about30 mV to about 90 mV, from about 30 mV to about 80 mV, from about 30 mVto about 70 mV, from about 30 mV to about 60 mV, from about 30 mV toabout 50 mV, from about 30 mV to about 40 mV, from about 40 mV to about100 mV, from about 40 mV to about 90 mV, from about 40 mV to about 80mV, from about 40 mV to about 70 mV, from about 40 mV to about 60 mV,and from about 40 mV to about 50 mV. In some embodiments, the zetapotential of the lipid nanoparticles may be from about 10 mV to about 50mV, from about 15 mV to about 45 mV, from about 20 mV to about 40 mV,and from about 25 mV to about 35 mV. In some embodiments, the zetapotential of the lipid nanoparticles may be about 10 mV, about 20 mV,about 30 mV, about 40 mV, about 50 mV, about 60 mV, about 70 mV, about80 mV, about 90 mV, and about 100 mV.

The term “encapsulation efficiency” of a polynucleotide describes theamount of the polynucleotide that is encapsulated by or otherwiseassociated with a nanoparticle composition after preparation, relativeto the initial amount provided. As used herein, “encapsulation” mayrefer to complete, substantial, or partial enclosure, confinement,surrounding, or encasement.

Encapsulation efficiency is desirably high (e.g., close to 100%). Theencapsulation efficiency may be measured, for example, by comparing theamount of the polynucleotide in a solution containing the nanoparticlecomposition before and after breaking up the nanoparticle compositionwith one or more organic solvents or detergents.

Fluorescence may be used to measure the amount of free polynucleotide ina solution. For the nanoparticle compositions described herein, theencapsulation efficiency of a polynucleotide may be at least 50%, forexample 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, 99%, or 100%. In some embodiments, the encapsulationefficiency may be at least 80%. In certain embodiments, theencapsulation efficiency may be at least 90%. The amount of apolynucleotide present in a pharmaceutical composition disclosed hereincan depend on multiple factors such as the size of the polynucleotide,desired target and/or application, or other properties of thenanoparticle composition as well as on the properties of thepolynucleotide.

For example, the amount of an mRNA useful in a nanoparticle compositionmay depend on the size (expressed as length, or molecular mass),sequence, and other characteristics of the mRNA. The relative amounts ofa polynucleotide in a nanoparticle composition may also vary.

The relative amounts of the lipid composition and the polynucleotidepresent in a lipid nanoparticle composition of the present disclosurecan be optimized according to considerations of efficacy andtolerability. For compositions including an mRNA as a polynucleotide,the N:P ratio can serve as a useful metric.

As the N:P ratio of a nanoparticle composition controls both expressionand tolerability, nanoparticle compositions with low N:P ratios andstrong expression are desirable. N:P ratios vary according to the ratioof lipids to RNA in a nanoparticle composition.

In general, a lower N:P ratio is preferred. The one or more RNA, lipids,and amounts thereof may be selected to provide an N:P ratio from about2:1 to about 30:1, such as 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1,12:1, 14:1, 16:1, 18:1, 20:1, 22:1, 24:1, 26:1, 28:1, or 30:1. Incertain embodiments, the N:P ratio may be from about 2:1 to about 8:1.In other embodiments, the N:P ratio is from about 5:1 to about 8:1. Incertain embodiments, the N:P ratio is between 5:1 and 6:1. In onespecific aspect, the N:P ratio is about is about 5.67:1.

In addition to providing nanoparticle compositions, the presentdisclosure also provides methods of producing lipid nanoparticlescomprising encapsulating a polynucleotide. Such method comprises usingany of the pharmaceutical compositions disclosed herein and producinglipid nanoparticles in accordance with methods of production of lipidnanoparticles known in the art. See, e.g., Wang et al. (2015) “Deliveryof oligonucleotides with lipid nanoparticles” Adv. Drug Deliv. Rev.87:68-80; Silva et al. (2015) “Delivery Systems for Biopharmaceuticals.Part I: Nanoparticles and Microparticles” Curr. Pharm. Technol. 16:940-954; Naseri et al. (2015) “Solid Lipid Nanoparticles andNanostructured Lipid Carriers: Structure, Preparation and Application”Adv. Pharm. Bull. 5:305-13; Silva et al. (2015) “Lipid nanoparticles forthe delivery of biopharmaceuticals” Curr. Pharm. Biotechnol. 16:291-302,and references cited therein.

26. Other Delivery Agents

a. Liposomes, Lipoplexes, and Lipid Nanoparticles

In some embodiments, the compositions or formulations of the presentdisclosure comprise a delivery agent, e.g., a liposome, a lioplexes, alipid nanoparticle, or any combination thereof. The polynucleotidesdescribed herein can be formulated using one or more liposomes,lipoplexes, or lipid nanoparticles. Liposomes, lipoplexes, or lipidnanoparticles can be used to improve the efficacy of the polynucleotidesdirected protein production as these formulations can increase celltransfection by the polynucleotide; and/or increase the translation ofencoded protein. The liposomes, lipoplexes, or lipid nanoparticles canalso be used to increase the stability of the polynucleotides.

Liposomes are artificially-prepared vesicles that may primarily becomposed of a lipid bilayer and may be used as a delivery vehicle forthe administration of pharmaceutical formulations. Liposomes can be ofdifferent sizes. A multilamellar vesicle (MLV) may be hundreds ofnanometers in diameter, and may contain a series of concentric bilayersseparated by narrow aqueous compartments. A small unicellular vesicle(SUV) may be smaller than 50 nm in diameter, and a large unilamellarvesicle (LUV) may be between 50 and 500 nm in diameter. Liposome designmay include, but is not limited to, opsonins or ligands to improve theattachment of liposomes to unhealthy tissue or to activate events suchas, but not limited to, endocytosis. Liposomes may contain a low or ahigh pH value in order to improve the delivery of the pharmaceuticalformulations.

The formation of liposomes may depend on the pharmaceutical formulationentrapped and the liposomal ingredients, the nature of the medium inwhich the lipid vesicles are dispersed, the effective concentration ofthe entrapped substance and its potential toxicity, any additionalprocesses involved during the application and/or delivery of thevesicles, the optimal size, polydispersity and the shelf-life of thevesicles for the intended application, and the batch-to-batchreproducibility and scale up production of safe and efficient liposomalproducts, etc.

As a non-limiting example, liposomes such as synthetic membrane vesiclesmay be prepared by the methods, apparatus and devices described in U.S.Pub. Nos. US20130177638, US20130177637, US20130177636, US20130177635,US20130177634, US20130177633, US20130183375, US20130183373, andUS20130183372. In some embodiments, the polynucleotides described hereinmay be encapsulated by the liposome and/or it may be contained in anaqueous core that may then be encapsulated by the liposome as describedin, e.g., Intl. Pub. Nos. WO2012031046, WO2012031043, WO2012030901,WO2012006378, and WO2013086526; and U.S. Pub. Nos. US20130189351,US20130195969 and US20130202684. Each of the references in hereinincorporated by reference in its entirety.

In some embodiments, the polynucleotides described herein can beformulated in a cationic oil-in-water emulsion where the emulsionparticle comprises an oil core and a cationic lipid that can interactwith the polynucleotide anchoring the molecule to the emulsion particle.In some embodiments, the polynucleotides described herein can beformulated in a water-in-oil emulsion comprising a continuoushydrophobic phase in which the hydrophilic phase is dispersed. Exemplaryemulsions can be made by the methods described in Intl. Pub. Nos.WO2012006380 and WO201087791, each of which is herein incorporated byreference in its entirety.

In some embodiments, the polynucleotides described herein can beformulated in a lipid-polycation complex. The formation of thelipid-polycation complex can be accomplished by methods as described in,e.g., U.S. Pub. No. US20120178702. As a non-limiting example, thepolycation can include a cationic peptide or a polypeptide such as, butnot limited to, polylysine, polyornithine and/or polyarginine and thecationic peptides described in Intl. Pub. No. WO2012013326 or U.S. Pub.No. US20130142818. Each of the references is herein incorporated byreference in its entirety.

In some embodiments, the polynucleotides described herein can beformulated in a lipid nanoparticle (LNP) such as those described inIntl. Pub. Nos. WO2013123523, WO2012170930, WO2011127255 andWO2008103276; and U.S. Pub. No. US20130171646, each of which is hereinincorporated by reference in its entirety.

Lipid nanoparticle formulations typically comprise one or more lipids.In some embodiments, the lipid is a cationic or an ionizable lipid. Insome embodiments, lipid nanoparticle formulations further comprise othercomponents, including a phospholipid, a structural lipid, a quaternaryamine compound, and a molecule capable of reducing particle aggregation,for example a PEG or PEG-modified lipid.

Cationic and ionizable lipids may include those as described in, e.g.,Intl. Pub. Nos. WO2015199952, WO 2015130584, WO 2015011633, andWO2012040184 WO2013126803, WO2011153120, WO2011149733, WO2011090965,WO2011043913, WO2011022460, WO2012061259, WO2012054365, WO2012044638,WO2010080724, WO201021865, WO2008103276, and WO2013086373; U.S. Pat.Nos. 7,893,302, 7,404,969, 8,283,333, and 8,466,122; and U.S. Pub. Nos.US20110224447, US20120295832, US20150315112, US20100036115,US20120202871, US20130064894, US20130129785, US20130150625,US20130178541, US20130123338 and US20130225836, each of which is hereinincorporated by reference in its entirety. In some embodiments, theamount of the cationic and ionizable lipids in the lipid compositionranges from about 0.01 mol % to about 99 mol %.

Exemplary ionizable lipids include, but not limited to, any one ofCompounds 1-147 disclosed herein, DLin-MC3-DMA (MC3), DLin-DMA, DLenDMA,DLin-D-DMA, DLin-K-DMA, DLin-M-C2-DMA, DLin-K-DMA, DLin-KC2-DMA,DLin-KC3-DMA, DLin-KC4-DMA, DLin-C2K-DMA, DLin-MP-DMA, DODMA, 98N12-5,C12-200, DLin-C-DAP, DLin-DAC, DLinDAP, DLinAP, DLin-EG-DMA,DLin-2-DMAP, KL10, KL22, KL25, Octyl-CLinDMA, Octyl-CLinDMA (2R),Octyl-CLinDMA (2S), and any combination thereof. Other exemplaryionizable lipids include,(13Z,16Z)-N,N-dimethyl-3-nonyldocosa-13,16-dien-1-amine (L608),(20Z,23Z)-N,N-dimethylnonacosa-20,23-dien-10-amine,(17Z,20Z)-N,N-dimemylhexacosa-17,20-dien-9-amine,(16Z,19Z)-N5N-dimethylpentacosa-16,19-dien-8-amine,(13Z,16Z)-N,N-dimethyldocosa-13,16-dien-5-amine,(12Z,15Z)-N,N-dimethylhenicosa-12,15-dien-4-amine,(14Z,17Z)-N,N-dimethyltricosa-14,17-dien-6-amine,(15Z,18Z)-N,N-dimethyltetracosa-15,18-dien-7-amine,(18Z,21Z)-N,N-dimethylheptacosa-18,21-dien-10-amine,(15Z,18Z)-N,N-dimethyltetracosa-15,18-dien-5-amine,(14Z,17Z)-N,N-dimethyltricosa-14,17-dien-4-amine,(19Z,22Z)-N,N-dimeihyloctacosa-19,22-dien-9-amine,(18Z,21Z)-N,N-dimethylheptacosa-18,21-dien-8-amine,(17Z,20Z)-N,N-dimethylhexacosa-17,20-dien-7-amine,(16Z,19Z)-N,N-dimethylpentacosa-16,19-dien-6-amine,(22Z,25Z)-N,N-dimethylhentriaconta-22,25-dien-10-amine,(21Z,24Z)-N,N-dimethyltriaconta-21,24-dien-9-amine,(18Z)-N,N-dimetylheptacos-18-en-10-amine,(17Z)-N,N-dimethylhexacos-17-en-9-amine,(19Z,22Z)-N,N-dimethyloctacosa-19,22-dien-7-amine,N,N-dimethylheptacosan-10-amine,(20Z,23Z)-N-ethyl-N-methylnonacosa-20,23-dien-10-amine,1-[(11Z,14Z)-1-nonylicosa-11,14-dien-1-yl]pyrrolidine,(20Z)-N,N-dimethylheptacos-20-en-10-amine, (15Z)-N,N-dimethyleptacos-15-en-10-amine, (14Z)-N,N-dimethylnonacos-14-en-10-amine,(17Z)-N,N-dimethylnonacos-17-en-10-amine,(24Z)-N,N-dimethyltritriacont-24-en-10-amine,(20Z)-N,N-dimethylnonacos-20-en-10-amine,(22Z)-N,N-dimethylhentriacont-22-en-10-amine,(16Z)-N,N-dimethylpentacos-16-en-8-amine,(12Z,15Z)-N,N-dimethyl-2-nonylhenicosa-12,15-dien-1-amine,N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl] eptadecan-8-amine,1-[(1S,2R)-2-hexylcyclopropyl]-N,N-dimethylnonadecan-10-amine,N,N-dimethyl-1-[(1 S,2R)-2-octylcyclopropyl]nonadecan amine,N,N-dimethyl-21-[(1S,2R)-2-octylcyclopropyl]henicosan-10-amine,N,N-dimethyl-1-[(1S,2S)-2-{[(1R,2R)-2-pentylcycIopropyl]methyl}cyclopropyl]nonadecan-10-amine,N,N-dimethyl [(1S,2R)-2-octylcyclopropyl]hexadecan-8-amine,N,N-dimethyl--[(1R,2S) undecyIcyclopropyl]tetradecan-5-amine,N,N-dimethyl-3-{7-[(1S,2R) octylcyclopropyl]heptyl}dodecan-1-amine,1-[(1R,2S)-2-heptylcyclopropyl]-N,N-dimethyloctadecan-9-amine,1-[(1S,2R)-2-decylcyclopropyl]-N,N-dimethylpentadecan-6-amine,N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]pentadecan-8-amine,R—N,N-dimethyl-1-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-3-(octyloxy)propan-2-amine,S—N,N-dimethyl-1-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-3-(octyloxy)propan-2-amine,1-{2-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-1-[(octyloxy)methyl]ethyl}pyrrolidine,(2S)—N,N-dimethyl-1-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-3-[(5Z)-oct-5-en-1-yloxy]propan-2-amine,1-{2-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-1-[(octyloxy)methyl]ethyl}azetidine,(2S)-1-(hexyloxy)-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]propan-2-amine,(2S)-1-(heptyloxy)-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]propan-2-amine,N,N-dimethyl-1-(nonyloxy)-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]propan-2-amine,N,N-dimethyl-1-[(9Z)-octadec-9-en-1-yloxy]-3-(octyloxy)propan-2-amine;(2S)—N,N-dimethyl-1-[(6Z,9Z,12Z)-octadeca-6,9,12-trien-1-yloxy]-3-(octyloxy)propan-2-amine,(2S)-1-[(11Z,14Z)-icosa-11,14-dien-1-yloxy]-N,N-dimethyl-3-(pentyloxy)propan-2-amine,(2S)-1-(hexyloxy)-3-[(11Z,14Z)-icosa-11,14-dien-1-yloxy]-N,N-dimethylpropan-2-amine,1-[(11Z,14Z)-icosa-11,14-dien-1-yloxy]-N,N-dimethyl-3-(octyloxy)propan-2-amine,1-[(13Z,16Z)-docosa-13,16-dien-1-yloxy]-N,N-dimethyl-3-(octyloxy)propan-2-amine,(2S)-1-[(13Z,16Z)-docosa-13,16-dien-1-yloxy]-3-(hexyloxy)-N,N-dimethylpropan-2-amine,(2S)-1-[(13Z)-docos-13-en-1-yloxy]-3-(hexyloxy)-N,N-dimethylpropan-2-amine,1-[(13Z)-docos-13-en-1-yloxy]-N,N-dimethyl-3-(octyloxy)propan-2-amine,1-[(9Z)-hexadec-9-en-1-yloxy]-N,N-dimethyl-3-(octyloxy)propan-2-amine,(2R)—N,N-dimethyl-H(1-metoyloctyl)oxy]-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]propan-2-amine,(2R)-1-[(3,7-dimethyloctyl)oxy]-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]propan-2-amine,N,N-dimethyl-1-(octyloxy)-3-({8-[(1S,2S)-2-{[(1R,2R)-2-pentylcyclopropyl]methyl}cyclopropyl]octyl}oxy)propan-2-amine,N,N-dimethyl-1-{[8-(2-oclylcyclopropyl)octyl]oxy}-3-(octyloxy)propan-2-amine,and (11E,20Z,23Z)-N,N-dimethylnonacosa-11,20,2-trien-10-amine, and anycombination thereof.

Phospholipids include, but are not limited to, glycerophospholipids suchas phosphatidylcholines, phosphatidylethanolamines, phosphatidylserines,phosphatidylinositols, phosphatidy glycerols, and phosphatidic acids.Phospholipids also include phosphosphingolipid, such as sphingomyelin.In some embodiments, the phospholipids are DLPC, DMPC, DOPC, DPPC, DSPC,DUPC, 18:0 Diether PC, DLnPC, DAPC, DHAPC, DOPE, 4ME 16:0 PE, DSPE,DLPE, DLnPE, DAPE, DHAPE, DOPG, and any combination thereof. In someembodiments, the phospholipids are MPPC, MSPC, PMPC, PSPC, SMPC, SPPC,DHAPE, DOPG, and any combination thereof. In some embodiments, theamount of phospholipids (e.g., DSPC and/or MSPC) in the lipidcomposition ranges from about 1 mol % to about 20 mol %.

The structural lipids include sterols and lipids containing sterolmoieties. In some embodiments, the structural lipids includecholesterol, fecosterol, sitosterol, ergosterol, campesterol,stigmasterol, brassicasterol, tomatidine, tomatine, ursolic acid,alpha-tocopherol, and mixtures thereof. In some embodiments, thestructural lipid is cholesterol. In some embodiments, the amount of thestructural lipids (e.g., cholesterol) in the lipid composition rangesfrom about 20 mol % to about 60 mol %.

The quaternary amine compound as described herein include1,2-dioleoyl-3-trimethylammonium-propane (DOTAP),N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA),1-[2-(oleoyloxy)ethyl]-2-oleyl-3-(2-hydroxyethyl)imidazolinium chloride(DOTIM),2,3-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-N,N-dimethyl-1-propanaminiumtrifluoroacetate (DOSPA), N,N-distearyl-N,N-dimethylammonium bromide(DDAB), N-(1,2-dimyristyloxyprop-3-yl)-N,N-dimethyl-N-hydroxyethylammonium bromide (DMRIE),N-(1,2-dioleoyloxyprop-3-yl)-N,N-dimethyl-N-hydroxyethyl ammoniumbromide (DOME), N,N-dioleyl-N,N-dimethylammonium chloride (DODAC),1,2-dilauroyl-sn-glycero-3-ethylphosphocholine (DLePC), 1,2-distearoyl-3-trimethylammonium-propane (DSTAP),1,2-dipalmitoyl-3-trimethylammonium-propane (DPTAP),1,2-dilinoleoyl-3-trimethylammonium-propane (DLTAP),1,2-dimyristoyl-3-trimethylammonium-propane (DMTAP), 1,2-distearoyl-sn-glycero-3-ethylphosphocholine (DSePC),1,2-dipalmitoyl-sn-glycero-3-ethylphosphocholine (DPePC), 1,2-dimyristoyl-sn-glycero-3-ethylphosphocholine (DMePC),1,2-dioleoyl-sn-glycero-3-ethylphosphocholine (DOePC),1,2-di-(9Z-tetradecenoyl)-sn-glycero-3-ethylphosphocholine (14:1 EPC),1-palmitoyl-2-oleoyl-sn-glycero-3-ethylphosphocholine (16:0-18:1 EPC),and any combination thereof. In some embodiments, the amount of thequaternary amine compounds (e.g., DOTAP) in the lipid composition rangesfrom about 0.01 mol % to about 20 mol %.

The PEG-modified lipids include PEG-modified phosphatidylethanolamineand phosphatidic acid, PEG-ceramide conjugates (e.g., PEG-CerC14 orPEG-CerC20), PEG-modified dialkylamines and PEG-modified1,2-diacyloxypropan-3-amines. Such lipids are also referred to asPEGylated lipids. For example, a PEG lipid may be PEG-c-DOMG, PEG-DMG,PEG-DLPE, PEG DMPE, PEG-DPPC, or a PEG-DSPE lipid. In some embodiments,the PEG-lipid are 1,2-dimyristoyl-sn-glycerol methoxypolyethylene glycol(PEG-DMG),1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethyleneglycol)] (PEG-DSPE), PEG-disteryl glycerol (PEG-DSG), PEG-dipalmetoleyl,PEG-dioleyl, PEG-distearyl, PEG-diacylglycamide (PEG-DAG),PEG-dipalmitoyl phosphatidylethanolamine (PEG-DPPE), orPEG-1,2-dimyristyloxlpropyl-3-amine (PEG-c-DMA). In some embodiments,the PEG moiety has a size of about 1000, 2000, 5000, 10,000, 15,000 or20,000 daltons. In some embodiments, the amount of PEG-lipid in thelipid composition ranges from about 0.1 mol % to about 5 mol %.

In some embodiments, the LNP formulations described herein canadditionally comprise a permeability enhancer molecule. Non-limitingpermeability enhancer molecules are described in U.S. Pub. No.US20050222064, herein incorporated by reference in its entirety.

The LNP formulations can further contain a phosphate conjugate. Thephosphate conjugate can increase in vivo circulation times and/orincrease the targeted delivery of the nanoparticle. Phosphate conjugatescan be made by the methods described in, e.g., Intl. Pub. No.WO2013033438 or U.S. Pub. No. US20130196948. The LNP formulation mayalso contain a polymer conjugate (e.g., a water soluble conjugate) asdescribed in, e.g., U.S. Pub. Nos. US20130059360, US20130196948, andUS20130072709. Each of the references is herein incorporated byreference in its entirety.

The LNP formulations can comprise a conjugate to enhance the delivery ofnanoparticles of the present disclosure in a subject. Further, theconjugate can inhibit phagocytic clearance of the nanoparticles in asubject. In some embodiments, the conjugate may be a “self” peptidedesigned from the human membrane protein CD47 (e.g., the “self”particles described by Rodriguez et al, Science 2013 339, 971-975,herein incorporated by reference in its entirety). As shown by Rodriguezet al. the self-peptides delayed macrophage-mediated clearance ofnanoparticles which enhanced delivery of the nanoparticles.

The LNP formulations can comprise a carbohydrate carrier. As anon-limiting example, the carbohydrate carrier can include, but is notlimited to, an anhydride-modified phytoglycogen or glycogen-typematerial, phytoglycogen octenyl succinate, phytoglycogen beta-dextrin,anhydride-modified phytoglycogen beta-dextrin (e.g., Intl. Pub. No.WO2012109121, herein incorporated by reference in its entirety).

The LNP formulations can be coated with a surfactant or polymer toimprove the delivery of the particle. In some embodiments, the LNP maybe coated with a hydrophilic coating such as, but not limited to, PEGcoatings and/or coatings that have a neutral surface charge as describedin U.S. Pub. No. US20130183244, herein incorporated by reference in itsentirety.

The LNP formulations can be engineered to alter the surface propertiesof particles so that the lipid nanoparticles may penetrate the mucosalbarrier as described in U.S. Pat. No. 8,241,670 or Intl. Pub. No.WO2013110028, each of which is herein incorporated by reference in itsentirety.

The LNP engineered to penetrate mucus can comprise a polymeric material(i.e., a polymeric core) and/or a polymer-vitamin conjugate and/or atri-block co-polymer. The polymeric material can include, but is notlimited to, polyamines, polyethers, polyamides, polyesters,polycarbamates, polyureas, polycarbonates, poly(styrenes), polyimides,polysulfones, polyurethanes, polyacetylenes, polyethylenes,polyethyeneimines, polyisocyanates, polyacrylates, polymethacrylates,polyacrylonitriles, and polyarylates.

LNP engineered to penetrate mucus can also include surface alteringagents such as, but not limited to, polynucleotides, anionic proteins(e.g., bovine serum albumin), surfactants (e.g., cationic surfactantssuch as for example dimethyldioctadecyl-ammonium bromide), sugars orsugar derivatives (e.g., cyclodextrin), nucleic acids, polymers (e.g.,heparin, polyethylene glycol and poloxamer), mucolytic agents (e.g.,N-acetylcysteine, mugwort, bromelain, papain, clerodendrum,acetylcysteine, bromhexine, carbocisteine, eprazinone, mesna, ambroxol,sobrerol, domiodol, letosteine, stepronin, tiopronin, gelsolin, thymosinβ4 dornase alfa, neltenexine, erdosteine) and various DNases includingrhDNase.

In some embodiments, the mucus penetrating LNP can be a hypotonicformulation comprising a mucosal penetration enhancing coating. Theformulation can be hypotonic for the epithelium to which it is beingdelivered. Non-limiting examples of hypotonic formulations may be foundin, e.g., Intl. Pub. No. WO2013110028, herein incorporated by referencein its entirety.

In some embodiments, the polynucleotide described herein is formulatedas a lipoplex, such as, without limitation, the ATUPLEX™ system, theDACC system, the DBTC system and other siRNA-lipoplex technology fromSilence Therapeutics (London, United Kingdom), STEMFECT™ from STEMGENT®(Cambridge, Mass.), and polyethylenimine (PEI) or protamine-basedtargeted and non-targeted delivery of nucleic acids (Aleku et al. CancerRes. 2008 68:9788-9798; Strumberg et al. Int J Clin Pharmacol Ther 201250:76-78; Santel et al., Gene Ther 2006 13:1222-1234; Santel et al.,Gene Ther 2006 13:1360-1370; Gutbier et al., Pulm Pharmacol. Ther. 201023:334-344; Kaufmann et al. Microvasc Res 2010 80:286-293Weide et al. JImmunother. 2009 32:498-507; Weide et al. J Immunother. 2008 31:180-188;Pascolo Expert Opin. Biol. Ther. 4:1285-1294; Fotin-Mleczek et al., 2011J. Immunother. 34:1-15; Song et al., Nature Biotechnol. 2005,23:709-717; Peer et al., Proc Natl Acad Sci USA. 2007 6; 104:4095-4100;deFougerolles Hum Gene Ther. 2008 19:125-132; all of which areincorporated herein by reference in its entirety).

In some embodiments, the polynucleotides described herein are formulatedas a solid lipid nanoparticle (SLN), which may be spherical with anaverage diameter between 10 to 1000 nm. SLN possess a solid lipid corematrix that can solubilize lipophilic molecules and may be stabilizedwith surfactants and/or emulsifiers. Exemplary SLN can be those asdescribed in Intl. Pub. No. WO2013105101, herein incorporated byreference in its entirety.

In some embodiments, the polynucleotides described herein can beformulated for controlled release and/or targeted delivery. As usedherein, “controlled release” refers to a pharmaceutical composition orcompound release profile that conforms to a particular pattern ofrelease to effect a therapeutic outcome. In one embodiment, thepolynucleotides can be encapsulated into a delivery agent describedherein and/or known in the art for controlled release and/or targeteddelivery. As used herein, the term “encapsulate” means to enclose,surround or encase. As it relates to the formulation of the compounds ofthe disclosure, encapsulation may be substantial, complete or partial.The term “substantially encapsulated” means that at least greater than50, 60, 70, 80, 85, 90, 95, 96, 97, 98, 99, 99.9, 99.9 or greater than99.999% of the pharmaceutical composition or compound of the disclosuremay be enclosed, surrounded or encased within the delivery agent.“Partially encapsulation” means that less than 10, 10, 20, 30, 40 50 orless of the pharmaceutical composition or compound of the disclosure maybe enclosed, surrounded or encased within the delivery agent.

Advantageously, encapsulation can be determined by measuring the escapeor the activity of the pharmaceutical composition or compound of thedisclosure using fluorescence and/or electron micrograph. For example,at least 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 85, 90, 95, 96, 97, 98,99, 99.9, 99.99 or greater than 99.99% of the pharmaceutical compositionor compound of the disclosure are encapsulated in the delivery agent.

In some embodiments, the polynucleotide controlled release formulationcan include at least one controlled release coating (e.g., OPADRY®,EUDRAGIT RL®, EUDRAGIT RS® and cellulose derivatives such asethylcellulose aqueous dispersions (AQUACOAT® and SURELEASE®)). In someembodiments, the polynucleotide controlled release formulation cancomprise a polymer system as described in U.S. Pub. No. US20130130348,or a PEG and/or PEG related polymer derivative as described in U.S. Pat.No. 8,404,222, each of which is incorporated by reference in itsentirety.

In some embodiments, the polynucleotides described herein can beencapsulated in a therapeutic nanoparticle, referred to herein as“therapeutic nanoparticle polynucleotides.” Therapeutic nanoparticlesmay be formulated by methods described in, e.g., Intl. Pub. Nos.WO2010005740, WO2010030763, WO2010005721, WO2010005723, andWO2012054923; and U.S. Pub. Nos. US20110262491, US20100104645,US20100087337, US20100068285, US20110274759, US20100068286,US20120288541, US20120140790, US20130123351 and US20130230567; and U.S.Pat. Nos. 8,206,747, 8,293,276, 8,318,208 and 8,318,211, each of whichis herein incorporated by reference in its entirety.

In some embodiments, the therapeutic nanoparticle polynucleotide can beformulated for sustained release. As used herein, “sustained release”refers to a pharmaceutical composition or compound that conforms to arelease rate over a specific period of time. The period of time mayinclude, but is not limited to, hours, days, weeks, months and years. Asa non-limiting example, the sustained release nanoparticle of thepolynucleotides described herein can be formulated as disclosed in Intl.Pub. No. WO2010075072 and U.S. Pub. Nos. US20100216804, US20110217377,US20120201859 and US20130150295, each of which is herein incorporated byreference in their entirety.

In some embodiments, the therapeutic nanoparticle polynucleotide can beformulated to be target specific, such as those described in Intl. Pub.Nos. WO2008121949, WO2010005726, WO2010005725, WO2011084521 andWO2011084518; and U.S. Pub. Nos. US20100069426, US20120004293 andUS20100104655, each of which is herein incorporated by reference in itsentirety.

The LNPs can be prepared using microfluidic mixers or micromixers.Exemplary microfluidic mixers can include, but are not limited to, aslit interdigitial micromixer including, but not limited to thosemanufactured by Microinnova (Allerheiligen bei Wildon, Austria) and/or astaggered herringbone micromixer (SHM) (see Zhigaltsevet al., “Bottom-updesign and synthesis of limit size lipid nanoparticle systems withaqueous and triglyceride cores using millisecond microfluidic mixing,”Langmuir 28:3633-40 (2012); Belliveau et al., “Microfluidic synthesis ofhighly potent limit-size lipid nanoparticles for in vivo delivery ofsiRNA,” Molecular Therapy-Nucleic Acids. 1:e37 (2012); Chen et al.,“Rapid discovery of potent siRNA-containing lipid nanoparticles enabledby controlled microfluidic formulation,” J. Am. Chem. Soc.134(16):6948-51 (2012); each of which is herein incorporated byreference in its entirety). Exemplary micromixers include SlitInterdigital Microstructured Mixer (SIMM-V2) or a Standard SlitInterdigital Micro Mixer (SSIMM) or Caterpillar (CPMM) or Impinging-jet(IJMM,) from the Institut für Mikrotechnik Mainz GmbH, Mainz Germany. Insome embodiments, methods of making LNP using SHM further comprisemixing at least two input streams wherein mixing occurs bymicrostructure-induced chaotic advection (MICA). According to thismethod, fluid streams flow through channels present in a herringbonepattern causing rotational flow and folding the fluids around eachother. This method can also comprise a surface for fluid mixing whereinthe surface changes orientations during fluid cycling. Methods ofgenerating LNPs using SHM include those disclosed in U.S. Pub. Nos.US20040262223 and US20120276209, each of which is incorporated herein byreference in their entirety.

In some embodiments, the polynucleotides described herein can beformulated in lipid nanoparticles using microfluidic technology (seeWhitesides, George M., “The Origins and the Future of Microfluidics,”Nature 442: 368-373 (2006); and Abraham et al., “Chaotic Mixer forMicrochannels,” Science 295: 647-651 (2002); each of which is hereinincorporated by reference in its entirety). In some embodiments, thepolynucleotides can be formulated in lipid nanoparticles using amicromixer chip such as, but not limited to, those from HarvardApparatus (Holliston, Mass.) or Dolomite Microfluidics (Royston, UK). Amicromixer chip can be used for rapid mixing of two or more fluidstreams with a split and recombine mechanism.

In some embodiments, the polynucleotides described herein can beformulated in lipid nanoparticles having a diameter from about 1 nm toabout 100 nm such as, but not limited to, about 1 nm to about 20 nm,from about 1 nm to about 30 nm, from about 1 nm to about 40 nm, fromabout 1 nm to about 50 nm, from about 1 nm to about 60 nm, from about 1nm to about 70 nm, from about 1 nm to about 80 nm, from about 1 nm toabout 90 nm, from about 5 nm to about from 100 nm, from about 5 nm toabout 10 nm, about 5 nm to about 20 nm, from about 5 nm to about 30 nm,from about 5 nm to about 40 nm, from about 5 nm to about 50 nm, fromabout 5 nm to about 60 nm, from about 5 nm to about 70 nm, from about 5nm to about 80 nm, from about 5 nm to about 90 nm, about 10 to about 20nm, about 10 to about 30 nm, about 10 to about 40 nm, about 10 to about50 nm, about 10 to about 60 nm, about 10 to about 70 nm, about 10 toabout 80 nm, about 10 to about 90 nm, about 20 to about 30 nm, about 20to about 40 nm, about 20 to about 50 nm, about 20 to about 60 nm, about20 to about 70 nm, about 20 to about 80 nm, about 20 to about 90 nm,about 20 to about 100 nm, about 30 to about 40 nm, about 30 to about 50nm, about 30 to about 60 nm, about 30 to about 70 nm, about 30 to about80 nm, about 30 to about 90 nm, about 30 to about 100 nm, about 40 toabout 50 nm, about 40 to about 60 nm, about 40 to about 70 nm, about 40to about 80 nm, about 40 to about 90 nm, about 40 to about 100 nm, about50 to about 60 nm, about 50 to about 70 nm about 50 to about 80 nm,about 50 to about 90 nm, about 50 to about 100 nm, about 60 to about 70nm, about 60 to about 80 nm, about 60 to about 90 nm, about 60 to about100 nm, about 70 to about 80 nm, about 70 to about 90 nm, about 70 toabout 100 nm, about 80 to about 90 nm, about 80 to about 100 nm and/orabout 90 to about 100 nm.

In some embodiments, the lipid nanoparticles can have a diameter fromabout 10 to 500 nm. In one embodiment, the lipid nanoparticle may have adiameter greater than 100 nm, greater than 150 nm, greater than 200 nm,greater than 250 nm, greater than 300 nm, greater than 350 nm, greaterthan 400 nm, greater than 450 nm, greater than 500 nm, greater than 550nm, greater than 600 nm, greater than 650 nm, greater than 700 nm,greater than 750 nm, greater than 800 nm, greater than 850 nm, greaterthan 900 nm, greater than 950 nm or greater than 1000 nm.

In some embodiments, the polynucleotides can be delivered using smallerLNPs. Such particles may comprise a diameter from below 0.1 μm up to 100nm such as, but not limited to, less than 0.1 μm, less than 1.0 μm, lessthan 5 μm, less than 10 μm, less than 15 um, less than 20 um, less than25 um, less than 30 um, less than 35 um, less than 40 um, less than 50um, less than 55 um, less than 60 um, less than 65 um, less than 70 um,less than 75 um, less than 80 um, less than 85 um, less than 90 um, lessthan 95 um, less than 100 um, less than 125 um, less than 150 um, lessthan 175 um, less than 200 um, less than 225 um, less than 250 um, lessthan 275 um, less than 300 um, less than 325 um, less than 350 um, lessthan 375 um, less than 400 um, less than 425 um, less than 450 um, lessthan 475 um, less than 500 um, less than 525 um, less than 550 um, lessthan 575 um, less than 600 um, less than 625 um, less than 650 um, lessthan 675 um, less than 700 um, less than 725 um, less than 750 um, lessthan 775 um, less than 800 um, less than 825 um, less than 850 um, lessthan 875 um, less than 900 um, less than 925 um, less than 950 um, orless than 975 um.

The nanoparticles and microparticles described herein can begeometrically engineered to modulate macrophage and/or the immuneresponse. The geometrically engineered particles can have varied shapes,sizes and/or surface charges to incorporate the polynucleotidesdescribed herein for targeted delivery such as, but not limited to,pulmonary delivery (see, e.g., Intl. Pub. No. WO2013082111, hereinincorporated by reference in its entirety). Other physical features thegeometrically engineering particles can include, but are not limited to,fenestrations, angled arms, asymmetry and surface roughness, charge thatcan alter the interactions with cells and tissues. In some embodiment,the nanoparticles described herein are stealth nanoparticles ortarget-specific stealth nanoparticles such as, but not limited to, thosedescribed in U.S. Pub. No. US20130172406, herein incorporated byreference in its entirety. The stealth or target-specific stealthnanoparticles can comprise a polymeric matrix, which may comprise two ormore polymers such as, but not limited to, polyethylenes,polycarbonates, polyanhydrides, polyhydroxyacids, polypropylfumerates,polycaprolactones, polyamides, polyacetals, polyethers, polyesters,poly(orthoesters), polycyanoacrylates, polyvinyl alcohols,polyurethanes, polyphosphazenes, polyacrylates, polymethacrylates,polycyanoacrylates, polyureas, polystyrenes, polyamines, polyesters,polyanhydrides, polyethers, polyurethanes, polymethacrylates,polyacrylates, polycyanoacrylates, or combinations thereof.

b. Lipidoids

In some embodiments, the compositions or formulations of the presentdisclosure comprise a delivery agent, e.g., a lipidoid. Thepolynucleotides described herein can be formulated with lipidoids.Complexes, micelles, liposomes or particles can be prepared containingthese lipidoids and therefore to achieve an effective delivery of thepolynucleotide, as judged by the production of an encoded protein,following the injection of a lipidoid formulation via localized and/orsystemic routes of administration. Lipidoid complexes of polynucleotidescan be administered by various means including, but not limited to,intravenous, intramuscular, or subcutaneous routes.

The synthesis of lipidoids is described in literature (see Mahon et al.,Bioconjug. Chem. 2010 21:1448-1454; Schroeder et al., J Intern Med. 2010267:9-21; Akinc et al., Nat Biotechnol. 2008 26:561-569; Love et al.,Proc Natl Acad Sci USA. 2010 107:1864-1869; Siegwart et al., Proc NatlAcad Sci USA. 2011 108:12996-3001; all of which are incorporated hereinin their entireties).

Formulations with the different lipidoids, including, but not limited topenta[3-(1-laurylaminopropionyl)]-triethylenetetramine hydrochloride(TETA-5LAP; also known as 98N12-5, see Murugaiah et al., AnalyticalBiochemistry, 401:61 (2010)), C12-200 (including derivatives andvariants), and MD1, can be tested for in vivo activity. The lipidoid“98N12-5” is disclosed by Akinc et al., Mol Ther. 2009 17:872-879. Thelipidoid “C12-200” is disclosed by Love et al., Proc Natl Acad Sci USA.2010 107:1864-1869 and Liu and Huang, Molecular Therapy. 2010 669-670.Each of the references is herein incorporated by reference in itsentirety.

In one embodiment, the polynucleotides described herein can beformulated in an aminoalcohol lipidoid. Aminoalcohol lipidoids may beprepared by the methods described in U.S. Pat. No. 8,450,298 (hereinincorporated by reference in its entirety).

The lipidoid formulations can include particles comprising either 3 or 4or more components in addition to polynucleotides. Lipidoids andpolynucleotide formulations comprising lipidoids are described in Intl.Pub. No. WO 2015051214 (herein incorporated by reference in itsentirety.

c. Hyaluronidase

In some embodiments, the polynucleotides described herein andhyaluronidase for injection (e.g., intramuscular or subcutaneousinjection). Hyaluronidase catalyzes the hydrolysis of hyaluronan, whichis a constituent of the interstitial barrier. Hyaluronidase lowers theviscosity of hyaluronan, thereby increases tissue permeability (Frost,Expert Opin. Drug Deliv. (2007) 4:427-440). Alternatively, thehyaluronidase can be used to increase the number of cells exposed to thepolynucleotides administered intramuscularly, intratumorally, orsubcutaneously.

d. Nanoparticle Mimics

In some embodiments, the polynucleotides described herein isencapsulated within and/or absorbed to a nanoparticle mimic. Ananoparticle mimic can mimic the delivery function organisms orparticles such as, but not limited to, pathogens, viruses, bacteria,fungus, parasites, prions and cells. As a non-limiting example, thepolynucleotides described herein can be encapsulated in a non-vironparticle that can mimic the delivery function of a virus (see e.g.,Intl. Pub. No. WO2012006376 and U.S. Pub. Nos. US20130171241 andUS20130195968, each of which is herein incorporated by reference in itsentirety).

e. Nanotubes

In some embodiments, the compositions or formulations of the presentdisclosure comprise the polynucleotides described herein attached orotherwise bound to (e.g., through steric, ionic, covalent and/or otherforces) at least one nanotube, such as, but not limited to, rosettenanotubes, rosette nanotubes having twin bases with a linker, carbonnanotubes and/or single-walled carbon nanotubes. Nanotubes and nanotubeformulations comprising a polynucleotide are described in, e.g., Intl.Pub. No. WO2014152211, herein incorporated by reference in its entirety.

f. Self-Assembled Nanoparticles, or Self-Assembled Macromolecules

In some embodiments, the compositions or formulations of the presentdisclosure comprise the polynucleotides described herein inself-assembled nanoparticles, or amphiphilic macromolecules (AMs) fordelivery. AMs comprise biocompatible amphiphilic polymers that have analkylated sugar backbone covalently linked to poly(ethylene glycol). Inaqueous solution, the AMs self-assemble to form micelles. Nucleic acidself-assembled nanoparticles are described in Intl. Appl. No.PCT/US2014/027077, and AMs and methods of forming AMs are described inU.S. Pub. No. US20130217753, each of which is herein incorporated byreference in its entirety.

g. Inorganic Nanoparticles, Semi-Conductive and Metallic Nanoparticles

In some embodiments, the compositions or formulations of the presentdisclosure comprise the polynucleotides described herein in inorganicnanoparticles, or water-dispersible nanoparticles comprising asemiconductive or metallic material. The inorganic nanoparticles caninclude, but are not limited to, clay substances that are waterswellable. The water-dispersible nanoparticles can be hydrophobic orhydrophilic nanoparticles. As a non-limiting example, the inorganic,semi-conductive and metallic nanoparticles are described in, e.g., U.S.Pat. Nos. 5,585,108 and 8,257,745; and U.S. Pub. Nos. US20120228565, US20120265001 and US 20120283503, each of which is herein incorporated byreference in their entirety.

h. Surgical Sealants: Gels and Hydrogels

In some embodiments, the compositions or formulations of the presentdisclosure comprise the polynucleotides described herein in a surgicalsealant. Surgical sealants such as gels and hydrogels are described inIntl. Appl. No. PCT/US2014/027077, herein incorporated by reference inits entirety.

i. Suspension Formulations

In some embodiments, the compositions or formulations of the presentdisclosure comprise the polynucleotides described herein in suspensions.In some embodiments, suspensions comprise a polynucleotide, waterimmiscible oil depots, surfactants and/or co-surfactants and/orco-solvents. Suspensions can be formed by first preparing an aqueoussolution of a polynucleotide and an oil-based phase comprising one ormore surfactants, and then mixing the two phases (aqueous andoil-based).

Exemplary oils for suspension formulations can include, but are notlimited to, sesame oil and Miglyol (comprising esters of saturatedcoconut and palmkernel oil-derived caprylic and capric fatty acids andglycerin or propylene glycol), corn oil, soybean oil, peanut oil,beeswax and/or palm seed oil. Exemplary surfactants can include, but arenot limited to Cremophor, polysorbate 20, polysorbate 80, polyethyleneglycol, transcutol, Capmul®, labrasol, isopropyl myristate, and/or Span80. In some embodiments, suspensions can comprise co-solvents including,but not limited to ethanol, glycerol and/or propylene glycol.

In some embodiments, suspensions can provide modulation of the releaseof the polynucleotides into the surrounding environment by diffusionfrom a water immiscible depot followed by resolubilization into asurrounding environment (e.g., an aqueous environment).

In some embodiments, the polynucleotides can be formulated such thatupon injection, an emulsion forms spontaneously (e.g., when delivered toan aqueous phase), which may provide a high surface area to volume ratiofor release of polynucleotides from an oil phase to an aqueous phase. Insome embodiments, the polynucleotide is formulated in a nanoemulsion,which can comprise a liquid hydrophobic core surrounded by or coatedwith a lipid or surfactant layer. Exemplary nanoemulsions and theirpreparations are described in, e.g., U.S. Pat. No. 8,496,945, hereinincorporated by reference in its entirety.

j. Cations and Anions

In some embodiments, the compositions or formulations of the presentdisclosure comprise the polynucleotides described herein and a cation oranion, such as Zn2+, Ca2+, Cu2+, Mg2+ and combinations thereof.Exemplary formulations can include polymers and a polynucleotidecomplexed with a metal cation as described in, e.g., U.S. Pat. Nos.6,265,389 and 6,555,525, each of which is herein incorporated byreference in its entirety. In some embodiments, cationic nanoparticlescan contain a combination of divalent and monovalent cations. Thedelivery of polynucleotides in cationic nanoparticles or in one or moredepot comprising cationic nanoparticles may improve polynucleotidebioavailability by acting as a long-acting depot and/or reducing therate of degradation by nucleases.

k. Molded Nanoparticles and Microparticles

In some embodiments, the compositions or formulations of the presentdisclosure comprise the polynucleotides described herein in moldednanoparticles in various sizes, shapes and chemistry. For example, thenanoparticles and/or microparticles can be made using the PRINT®technology by LIQUIDA TECHNOLOGIES® (Morrisville, N.C.) (e.g.,International Pub. No. WO2007024323, herein incorporated by reference inits entirety).

In some embodiments, the polynucleotides described herein is formulatedin microparticles. The microparticles may contain a core of thepolynucleotide and a cortex of a biocompatible and/or biodegradablepolymer, including but not limited to, poly(a-hydroxy acid), apolyhydroxy butyric acid, a polycaprolactone, a polyorthoester and apolyanhydride. The microparticle may have adsorbent surfaces to adsorbpolynucleotides. The microparticles may have a diameter of from at least1 micron to at least 100 microns (e.g., at least 1 micron, at least 10micron, at least 20 micron, at least 30 micron, at least 50 micron, atleast 75 micron, at least 95 micron, and at least 100 micron). In someembodiment, the compositions or formulations of the present disclosureare microemulsions comprising microparticles and polynucleotides.Exemplary microparticles, microemulsions and their preparations aredescribed in, e.g., U.S. Pat. Nos. 8,460,709, 8,309,139 and 8,206,749;U.S. Pub. Nos. US20130129830, US2013195923 and US20130195898; and Intl.Pub. No. WO2013075068, each of which is herein incorporated by referencein its entirety.

l. NanoJackets and NanoLiposomes

In some embodiments, the compositions or formulations of the presentdisclosure comprise the polynucleotides described herein in NanoJacketsand NanoLiposomes by Keystone Nano (State College, Pa.). NanoJackets aremade of materials that are naturally found in the body includingcalcium, phosphate and may also include a small amount of silicates.Nanojackets may have a size ranging from 5 to 50 nm.

NanoLiposomes are made of lipids such as, but not limited to, lipidsthat naturally occur in the body. NanoLiposomes may have a size rangingfrom 60-80 nm. In some embodiments, the polynucleotides disclosed hereinare formulated in a NanoLiposome such as, but not limited to, CeramideNanoLiposomes.

m. Cells or Minicells

In some embodiments, the compositions or formulations of the presentdisclosure comprise the polynucleotides described herein that istransfected ex vivo into cells, which are subsequently transplanted intoa subject. Cell-based formulations of the polynucleotide disclosedherein can be used to ensure cell transfection (e.g., in the cellularcarrier), alter the biodistribution of the polynucleotide (e.g., bytargeting the cell carrier to specific tissues or cell types), and/orincrease the translation of encoded protein.

Exemplary cells include, but are not limited to, red blood cells,virosomes, and electroporated cells (see e.g., Godfrin et al., ExpertOpin Biol Ther. 2012 12:127-133; Fang et al., Expert Opin Biol Ther.2012 12:385-389; Hu et al., Proc Natl Acad Sci USA. 2011108:10980-10985; Lund et al., Pharm Res. 2010 27:400-420; Huckriede etal., J Liposome Res. 2007; 17:39-47; Cusi, Hum Vaccin. 2006 2:1-7; deJonge et al., Gene Ther. 2006 13:400-411; all of which are hereinincorporated by reference in its entirety).

A variety of methods are known in the art and are suitable forintroduction of nucleic acid into a cell, including viral and non-viralmediated techniques. Examples of typical non-viral mediated techniquesinclude, but are not limited to, electroporation, calcium phosphatemediated transfer, nucleofection, sonoporation, heat shock,magnetofection, liposome mediated transfer, microinjection,microprojectile mediated transfer (nanoparticles), cationic polymermediated transfer (DEAE-dextran, polyethylenimine, polyethylene glycol(PEG) and the like) or cell fusion.

In some embodiments, the polynucleotides described herein can bedelivered in synthetic virus-like particles (VLPs) synthesized by themethods as described in Intl. Pub Nos. WO2011085231 and WO2013116656;and U.S. Pub. No. 20110171248, each of which is herein incorporated byreference in its entirety.

The technique of sonoporation, or cellular sonication, is the use ofsound (e.g., ultrasonic frequencies) for modifying the permeability ofthe cell plasma membrane. Sonoporation methods are known to delivernucleic acids in vivo (Yoon and Park, Expert Opin Drug Deliv. 20107:321-330; Postema and Gilja, Curr Pharm Biotechnol. 2007 8:355-361;Newman and Bettinger, Gene Ther. 2007 14:465-475; U.S. Pub. Nos.US20100196983 and US20100009424; all herein incorporated by reference intheir entirety).

In some embodiments, the polynucleotides described herein can bedelivered by electroporation. Electroporation techniques are known todeliver nucleic acids in vivo and clinically (Andre et al., Curr GeneTher. 2010 10:267-280; Chiarella et al., Curr Gene Ther. 201010:281-286; Hojman, Curr Gene Ther. 2010 10:128-138; all hereinincorporated by reference in their entirety). Electroporation devicesare sold by many companies worldwide including, but not limited to BTX®Instruments (Holliston, Mass.) (e.g., the AgilePulse In vivo System) andInovio (Blue Bell, Pa.) (e.g., Inovio SP-5P intramuscular deliverydevice or the CELLECTRA® 3000 intradermal delivery device).

In some embodiments, the cells are selected from the group consisting ofmammalian cells, bacterial cells, plant, microbial, algal and fungalcells. In some embodiments, the cells are mammalian cells, such as, butnot limited to, human, mouse, rat, goat, horse, rabbit, hamster or cowcells. In a further embodiment, the cells can be from an establishedcell line, including, but not limited to, HeLa, NSO, SP2/0, KEK 293T,Vero, Caco, Caco-2, MDCK, COS-1, COS-7, K562, Jurkat, CHO-K1, DG44,CHOK1SV, CHO-S, Huvec, CV-1, Huh-7, NIH3T3, HEK293, 293, A549, HepG2,IMR-90, MCF-7, U-20S, Per.C6, SF9, SF21 or Chinese Hamster Ovary (CHO)cells.

In certain embodiments, the cells are fungal cells, such as, but notlimited to, Chrysosporium cells, Aspergillus cells, Trichoderma cells,Dictyostelium cells, Candida cells, Saccharomyces cells,Schizosaccharomyces cells, and Penicillium cells.

In certain embodiments, the cells are bacterial cells such as, but notlimited to, E. coli, B. subtilis, or BL21 cells. Primary and secondarycells to be transfected by the methods of the disclosure can be obtainedfrom a variety of tissues and include, but are not limited to, all celltypes that can be maintained in culture. The primary and secondary cellsinclude, but are not limited to, fibroblasts, keratinocytes, epithelialcells (e.g., mammary epithelial cells, intestinal epithelial cells),endothelial cells, glial cells, neural cells, formed elements of theblood (e.g., lymphocytes, bone marrow cells), muscle cells andprecursors of these somatic cell types. Primary cells may also beobtained from a donor of the same species or from another species (e.g.,mouse, rat, rabbit, cat, dog, pig, cow, bird, sheep, goat, horse).

In some embodiments, the compositions or formulations of the presentdisclosure comprise the polynucleotides described herein in bacterialminicells. As a non-limiting example, bacterial minicells can be thosedescribed in Intl. Pub. No. WO2013088250 or U.S. Pub. No. US20130177499,each of which is herein incorporated by reference in its entirety.

n. Semi-Solid Compositions

In some embodiments, the compositions or formulations of the presentdisclosure comprise the polynucleotides described herein in ahydrophobic matrix to form a semi-solid or paste-like composition. As anon-limiting example, the semi-solid or paste-like composition can bemade by the methods described in Intl. Pub. No. WO201307604, hereinincorporated by reference in its entirety.

o. Exosomes

In some embodiments, the compositions or formulations of the presentdisclosure comprise the polynucleotides described herein in exosomes,which can be loaded with at least one polynucleotide and delivered tocells, tissues and/or organisms. As a non-limiting example, thepolynucleotides can be loaded in the exosomes as described in Intl. Pub.No. WO2013084000, herein incorporated by reference in its entirety.

p. Silk-Based Delivery

In some embodiments, the compositions or formulations of the presentdisclosure comprise the polynucleotides described herein that isformulated for silk-based delivery. The silk-based delivery system canbe formed by contacting a silk fibroin solution with a polynucleotidedescribed herein. As a non-limiting example, a sustained releasesilk-based delivery system and methods of making such system aredescribed in U.S. Pub. No. US20130177611, herein incorporated byreference in its entirety.

q. Amino Acid Lipids

In some embodiments, the compositions or formulations of the presentdisclosure comprise the polynucleotides described herein that isformulation with an amino acid lipid. Amino acid lipids are lipophiliccompounds comprising an amino acid residue and one or more lipophilictails. Non-limiting examples of amino acid lipids and methods of makingamino acid lipids are described in U.S. Pat. No. 8,501,824. The aminoacid lipid formulations may deliver a polynucleotide in releasable formthat comprises an amino acid lipid that binds and releases thepolynucleotides. As a non-limiting example, the release of thepolynucleotides described herein can be provided by an acid-labilelinker as described in, e.g., U.S. Pat. Nos. 7,098,032, 6,897,196,6,426,086, 7,138,382, 5,563,250, and 5,505,931, each of which is hereinincorporated by reference in its entirety.

r. Microvesicles

In some embodiments, the compositions or formulations of the presentdisclosure comprise the polynucleotides described herein in amicrovesicle formulation. Exemplary microvesicles include thosedescribed in U.S. Pub. No. US20130209544 (herein incorporated byreference in its entirety). In some embodiments, the microvesicle is anARRDC1-mediated microvesicles (ARMMs) as described in Intl. Pub. No.WO2013119602 (herein incorporated by reference in its entirety).

s. Interpolyelectrolyte Complexes

In some embodiments, the compositions or formulations of the presentdisclosure comprise the polynucleotides described herein in aninterpolyelectrolyte complex. Interpolyelectrolyte complexes are formedwhen charge-dynamic polymers are complexed with one or more anionicmolecules. Non-limiting examples of charge-dynamic polymers andinterpolyelectrolyte complexes and methods of makinginterpolyelectrolyte complexes are described in U.S. Pat. No. 8,524,368,herein incorporated by reference in its entirety.

t. Crystalline Polymeric Systems

In some embodiments, the compositions or formulations of the presentdisclosure comprise the polynucleotides described herein in crystallinepolymeric systems. Crystalline polymeric systems are polymers withcrystalline moieties and/or terminal units comprising crystallinemoieties. Exemplary polymers are described in U.S. Pat. No. 8,524,259(herein incorporated by reference in its entirety).

u. Polymers, Biodegradable Nanoparticles, and Core-Shell Nanoparticles

In some embodiments, the compositions or formulations of the presentdisclosure comprise the polynucleotides described herein and a naturaland/or synthetic polymer. The polymers include, but not limited to,polyethenes, polyethylene glycol (PEG), poly(l-lysine)(PLL), PEG graftedto PLL, cationic lipopolymer, biodegradable cationic lipopolymer,polyethyleneimine (PEI), cross-linked branched poly(alkylene imines), apolyamine derivative, a modified poloxamer, elastic biodegradablepolymer, biodegradable copolymer, biodegradable polyester copolymer,biodegradable polyester copolymer, multiblock copolymers, poly[α-(4-aminobutyl)-L-glycolic acid) (PAGA), biodegradable cross-linkedcationic multi-block copolymers, polycarbonates, polyanhydrides,polyhydroxyacids, polypropylfumerates, polycaprolactones, polyamides,polyacetals, polyethers, polyesters, poly(orthoesters),polycyanoacrylates, polyvinyl alcohols, polyurethanes, polyphosphazenes,polyureas, polystyrenes, polyamines, polylysine, poly(ethylene imine),poly(serine ester), poly(L-lactide-co-L-lysine),poly(4-hydroxy-L-proline ester), amine-containing polymers, dextranpolymers, dextran polymer derivatives or combinations thereof.

Exemplary polymers include, DYNAMIC POLYCONJUGATE® (Arrowhead ResearchCorp., Pasadena, Calif.) formulations from MIRUS® Bio (Madison, Wis.)and Roche Madison (Madison, Wis.), PHASERX™ polymer formulations suchas, without limitation, SMARTT POLYMER TECHNOLOGY™ (PHASERX®, Seattle,Wash.), DMRI/DOPE, poloxamer, VAXFECTIN® adjuvant from Vical (San Diego,Calif.), chitosan, cyclodextrin from Calando Pharmaceuticals (Pasadena,Calif.), dendrimers and poly(lactic-co-glycolic acid) (PLGA) polymers.RONDEL™ (RNAi/Oligonucleotide Nanoparticle Delivery) polymers (ArrowheadResearch Corporation, Pasadena, Calif.) and pH responsive co-blockpolymers such as PHASERX® (Seattle, Wash.).

The polymer formulations allow a sustained or delayed release of thepolynucleotide (e.g., following intramuscular or subcutaneousinjection). The altered release profile for the polynucleotide canresult in, for example, translation of an encoded protein over anextended period of time. The polymer formulation can also be used toincrease the stability of the polynucleotide. Sustained releaseformulations can include, but are not limited to, PLGA microspheres,ethylene vinyl acetate (EVAc), poloxamer, GELSITE® (Nanotherapeutics,Inc. Alachua, Fla.), HYLENEX® (Halozyme Therapeutics, San Diego Calif.),surgical sealants such as fibrinogen polymers (Ethicon Inc. Cornelia,Ga.), TISSELL® (Baxter International, Inc. Deerfield, Ill.), PEG-basedsealants, and COSEAL® (Baxter International, Inc. Deerfield, Ill.).

As a non-limiting example modified mRNA can be formulated in PLGAmicrospheres by preparing the PLGA microspheres with tunable releaserates (e.g., days and weeks) and encapsulating the modified mRNA in thePLGA microspheres while maintaining the integrity of the modified mRNAduring the encapsulation process. EVAc are non-biodegradable,biocompatible polymers that are used extensively in pre-clinicalsustained release implant applications (e.g., extended release productsOcusert a pilocarpine ophthalmic insert for glaucoma or progestasert asustained release progesterone intrauterine device; transdermal deliverysystems Testoderm, Duragesic and Selegiline; catheters). Poloxamer F-407NF is a hydrophilic, non-ionic surfactant triblock copolymer ofpolyoxyethylene-polyoxypropylene-polyoxyethylene having a low viscosityat temperatures less than 5° C. and forms a solid gel at temperaturesgreater than 15° C.

As a non-limiting example, the polynucleotides described herein can beformulated with the polymeric compound of PEG grafted with PLL asdescribed in U.S. Pat. No. 6,177,274. As another non-limiting example,the polynucleotides described herein can be formulated with a blockcopolymer such as a PLGA-PEG block copolymer (see e.g., U.S. Pub. No.US20120004293 and U.S. Pat. Nos. 8,236,330 and 8,246,968), or aPLGA-PEG-PLGA block copolymer (see e.g., U.S. Pat. No. 6,004,573). Eachof the references is herein incorporated by reference in its entirety.

In some embodiments, the polynucleotides described herein can beformulated with at least one amine-containing polymer such as, but notlimited to polylysine, polyethylene imine, poly(amidoamine) dendrimers,poly(amine-co-esters) or combinations thereof. Exemplary polyaminepolymers and their use as delivery agents are described in, e.g., U.S.Pat. Nos. 8,460,696, 8,236,280, each of which is herein incorporated byreference in its entirety.

In some embodiments, the polynucleotides described herein can beformulated in a biodegradable cationic lipopolymer, a biodegradablepolymer, or a biodegradable copolymer, a biodegradable polyestercopolymer, a biodegradable polyester polymer, a linear biodegradablecopolymer, PAGA, a biodegradable cross-linked cationic multi-blockcopolymer or combinations thereof as described in, e.g., U.S. Pat. Nos.6,696,038, 6,517,869, 6,267,987, 6,217,912, 6,652,886, 8,057,821, and8,444,992; U.S. Pub. Nos. US20030073619, US20040142474, US20100004315,US2012009145 and US20130195920; and Intl Pub. Nos. WO2006063249 andWO2013086322, each of which is herein incorporated by reference in itsentirety.

In some embodiments, the polynucleotides described herein can beformulated in or with at least one cyclodextrin polymer as described inU.S. Pub. No. US20130184453. In some embodiments, the polynucleotidesdescribed herein can be formulated in or with at least one crosslinkedcation-binding polymers as described in Intl. Pub. Nos. WO2013106072,WO2013106073 and WO2013106086. In some embodiments, the polynucleotidesdescribed herein can be formulated in or with at least PEGylated albuminpolymer as described in U.S. Pub. No. US20130231287. Each of thereferences is herein incorporated by reference in its entirety.

In some embodiments, the polynucleotides disclosed herein can beformulated as a nanoparticle using a combination of polymers, lipids,and/or other biodegradable agents, such as, but not limited to, calciumphosphate. Components can be combined in a core-shell, hybrid, and/orlayer-by-layer architecture, to allow for fine-tuning of thenanoparticle for delivery (Wang et al., Nat Mater. 2006 5:791-796;Fuller et al., Biomaterials. 2008 29:1526-1532; DeKoker et al., Adv DrugDeliv Rev. 2011 63:748-761; Endres et al., Biomaterials. 201132:7721-7731; Su et al., Mol Pharm. 2011 Jun. 6; 8(3):774-87; hereinincorporated by reference in their entireties). As a non-limitingexample, the nanoparticle can comprise a plurality of polymers such as,but not limited to hydrophilic-hydrophobic polymers (e.g., PEG-PLGA),hydrophobic polymers (e.g., PEG) and/or hydrophilic polymers (Intl. Pub.No. WO20120225129, herein incorporated by reference in its entirety).

The use of core-shell nanoparticles has additionally focused on ahigh-throughput approach to synthesize cationic cross-linked nanogelcores and various shells (Siegwart et al., Proc Natl Acad Sci USA. 2011108:12996-13001; herein incorporated by reference in its entirety). Thecomplexation, delivery, and internalization of the polymericnanoparticles can be precisely controlled by altering the chemicalcomposition in both the core and shell components of the nanoparticle.For example, the core-shell nanoparticles may efficiently deliver siRNAto mouse hepatocytes after they covalently attach cholesterol to thenanoparticle.

In some embodiments, a hollow lipid core comprising a middle PLGA layerand an outer neutral lipid layer containing PEG can be used to deliveryof the polynucleotides as described herein. In some embodiments, thelipid nanoparticles can comprise a core of the polynucleotides disclosedherein and a polymer shell, which is used to protect the polynucleotidesin the core. The polymer shell can be any of the polymers describedherein and are known in the art, the polymer shell can be used toprotect the polynucleotides in the core.

Core—shell nanoparticles for use with the polynucleotides describedherein are described in U.S. Pat. No. 8,313,777 or Intl. Pub. No.WO2013124867, each of which is herein incorporated by reference in theirentirety.

v. Peptides and Proteins

In some embodiments, the compositions or formulations of the presentdisclosure comprise the polynucleotides described herein that isformulated with peptides and/or proteins to increase transfection ofcells by the polynucleotide, and/or to alter the biodistribution of thepolynucleotide (e.g., by targeting specific tissues or cell types),and/or increase the translation of encoded protein (e.g., Intl. Pub.Nos. WO2012110636 and WO2013123298. In some embodiments, the peptidescan be those described in U.S. Pub. Nos. US20130129726, US20130137644and US20130164219. Each of the references is herein incorporated byreference in its entirety.

w. Conjugates

In some embodiments, the compositions or formulations of the presentdisclosure comprise the polynucleotides described herein that iscovalently linked to a carrier or targeting group, or including twoencoding regions that together produce a fusion protein (e.g., bearing atargeting group and therapeutic protein or peptide) as a conjugate. Theconjugate can be a peptide that selectively directs the nanoparticle toneurons in a tissue or organism, or assists in crossing the blood-brainbarrier.

The conjugates include a naturally occurring substance, such as aprotein (e.g., human serum albumin (HSA), low-density lipoprotein (LDL),high-density lipoprotein (HDL), or globulin); an carbohydrate (e.g., adextran, pullulan, chitin, chitosan, inulin, cyclodextrin or hyaluronicacid); or a lipid. The ligand may also be a recombinant or syntheticmolecule, such as a synthetic polymer, e.g., a synthetic polyamino acid,an oligonucleotide (e.g., an aptamer). Examples of polyamino acidsinclude polyamino acid is a polylysine (PLL), poly L-aspartic acid, polyL-glutamic acid, styrene-maleic acid anhydride copolymer,poly(L-lactide-co-glycolied) copolymer, divinyl ether-maleic anhydridecopolymer, N-(2-hydroxypropyl)methacrylamide copolymer (HMPA),polyethylene glycol (PEG), polyvinyl alcohol (PVA), polyurethane,poly(2-ethylacryllic acid), N-isopropylacrylamide polymers, orpolyphosphazine. Example of polyamines include: polyethylenimine,polylysine (PLL), spermine, spermidine, polyamine,pseudopeptide-polyamine, peptidomimetic polyamine, dendrimer polyamine,arginine, amidine, protamine, cationic lipid, cationic porphyrin,quaternary salt of a polyamine, or an alpha helical peptide.

In some embodiments, the conjugate can function as a carrier for thepolynucleotide disclosed herein. The conjugate can comprise a cationicpolymer such as, but not limited to, polyamine, polylysine,polyalkylenimine, and polyethylenimine that can be grafted to withpoly(ethylene glycol). Exemplary conjugates and their preparations aredescribed in U.S. Pat. No. 6,586,524 and U.S. Pub. No. US20130211249,each of which herein is incorporated by reference in its entirety.

The conjugates can also include targeting groups, e.g., a cell or tissuetargeting agent, e.g., a lectin, glycoprotein, lipid or protein, e.g.,an antibody, that binds to a specified cell type such as a kidney cell.A targeting group can be a thyrotropin, melanotropin, lectin,glycoprotein, surfactant protein A, Mucin carbohydrate, multivalentlactose, multivalent galactose, N-acetyl-galactosamine,N-acetyl-glucosamine multivalent mannose, multivalent fucose,glycosylated polyaminoacids, multivalent galactose, transferrin,bisphosphonate, polyglutamate, polyaspartate, a lipid, cholesterol, asteroid, bile acid, folate, vitamin B12, biotin, an RGD peptide, an RGDpeptide mimetic or an aptamer.

Targeting groups can be proteins, e.g., glycoproteins, or peptides,e.g., molecules having a specific affinity for a co-ligand, orantibodies e.g., an antibody, that binds to a specified cell type suchas a cancer cell, endothelial cell, or bone cell. Targeting groups mayalso include hormones and hormone receptors. They can also includenon-peptidic species, such as lipids, lectins, carbohydrates, vitamins,cofactors, multivalent lactose, multivalent galactose,N-acetyl-galactosamine, N-acetyl-glucosamine multivalent mannose,multivalent fructose, or aptamers. The ligand can be, for example, alipopolysaccharide, or an activator of p38 MAP kinase.

The targeting group can be any ligand that is capable of targeting aspecific receptor. Examples include, without limitation, folate, GalNAc,galactose, mannose, mannose-6P, apatamers, integrin receptor ligands,chemokine receptor ligands, transferrin, biotin, serotonin receptorligands, PSMA, endothelin, GCPII, somatostatin, LDL, and HDL ligands. Inparticular embodiments, the targeting group is an aptamer. The aptamercan be unmodified or have any combination of modifications disclosedherein. As a non-limiting example, the targeting group can be aglutathione receptor (GR)-binding conjugate for targeted delivery acrossthe blood-central nervous system barrier as described in, e.g., U.S.Pub. No. US2013021661012 (herein incorporated by reference in itsentirety).

In some embodiments, the conjugate can be a synergisticbiomolecule-polymer conjugate, which comprises a long-actingcontinuous-release system to provide a greater therapeutic efficacy. Thesynergistic biomolecule-polymer conjugate can be those described in U.S.Pub. No. US20130195799. In some embodiments, the conjugate can be anaptamer conjugate as described in Intl. Pat. Pub. No. WO2012040524. Insome embodiments, the conjugate can be an amine containing polymerconjugate as described in U.S. Pat. No. 8,507,653. Each of thereferences is herein incorporated by reference in its entirety. In someembodiments, the polynucleotides can be conjugated to SMARTT POLYMERTECHNOLOGY® (PHASERX®, Inc. Seattle, Wash.).

In some embodiments, the polynucleotides described herein are covalentlyconjugated to a cell penetrating polypeptide, which can also include asignal sequence or a targeting sequence. The conjugates can be designedto have increased stability, and/or increased cell transfection; and/oraltered the biodistribution (e.g., targeted to specific tissues or celltypes).

In some embodiments, the polynucleotides described herein can beconjugated to an agent to enhance delivery. In some embodiments, theagent can be a monomer or polymer such as a targeting monomer or apolymer having targeting blocks as described in Intl. Pub. No.WO2011062965. In some embodiments, the agent can be a transport agentcovalently coupled to a polynucleotide as described in, e.g., U.S. Pat.Nos. 6,835,393 and 7,374,778. In some embodiments, the agent can be amembrane barrier transport enhancing agent such as those described inU.S. Pat. Nos. 7,737,108 and 8,003,129. Each of the references is hereinincorporated by reference in its entirety.

x. Micro-Organs

In some embodiments, the compositions or formulations of the presentdisclosure comprise the polynucleotides described hereinin a micro-organthat can then express an encoded polypeptide of interest in along-lasting therapeutic formulation. Exemplary micro-organs andformulations are described in Intl. Pub. No. WO2014152211 (hereinincorporated by reference in its entirety).

y. Pseudovirions

In some embodiments, the compositions or formulations of the presentdisclosure comprise the polynucleotides described herein inpseudovirions (e.g., pseudovirions developed by Aura Biosciences,Cambridge, Mass.).

In some embodiments, the pseudovirion used for delivering thepolynucleotides can be derived from viruses such as, but not limited to,herpes and papillomaviruses as described in, e.g., U.S. Pub. Nos.US20130012450, US20130012566, US21030012426 and US20120207840; and Intl.Pub. No. WO2013009717, each of which is herein incorporated by referencein its entirety.

The pseudovirion can be a virus-like particle (VLP) prepared by themethods described in U.S. Pub. Nos. US20120015899 and US20130177587, andIntl. Pub. Nos. WO2010047839, WO2013116656, WO2013106525 andWO2013122262. In one aspect, the VLP can be bacteriophages MS, Qβ, R17,fr, GA, Sp, MI, I, MXI, NL95, AP205, f2, PP7, and the plant virusesTurnip crinkle virus (TCV), Tomato bushy stunt virus (TBSV), Southernbean mosaic virus (SBMV) and members of the genus Bromovirus includingBroad bean mottle virus, Brome mosaic virus, Cassia yellow blotch virus,Cowpea chlorotic mottle virus (CCMV), Melandrium yellow fleck virus, andSpring beauty latent virus. In another aspect, the VLP can be derivedfrom the influenza virus as described in U.S. Pub. No. US20130177587 andU.S. Pat. No. 8,506,967. In one aspect, the VLP can comprise a B7-1and/or B7-2 molecule anchored to a lipid membrane or the exterior of theparticle such as described in Intl. Pub. No. WO2013116656. In oneaspect, the VLP can be derived from norovirus, rotavirus recombinant VP6protein or double layered VP2/VP6 such as the VLP as described in Intl.Pub. No. WO2012049366. Each of the references is herein incorporated byreference in its entirety.

In some embodiments, the pseudovirion can be a human papillomavirus-like particle as described in Intl. Pub. No. WO2010120266 and U.S.Pub. No. US20120171290. In some embodiments, the virus-like particle(VLP) can be a self-assembled particle. In one aspect, the pseudovirionscan be virion derived nanoparticles as described in U.S. Pub. Nos.US20130116408 and US20130115247; and Intl. Pub. No. WO2013119877. Eachof the references is herein incorporated by reference in their entirety.

Non-limiting examples of formulations and methods for formulating thepolynucleotides described herein are also provided in Intl. Pub. NoWO2013090648 (incorporated herein by reference in their entirety).

27. Compositions and Formulations for Use

Certain aspects of the disclosure are directed to compositions orformulations comprising any of the polynucleotides disclosed above.

In some embodiments, the composition or formulation comprises:

-   -   (i) a polynucleotide (e.g., a RNA, e.g., an mRNA) comprising a        sequence-optimized nucleotide sequence encoding an IL-12        polypeptide as disclosed herein and/or a sequence-optimized        nucleotide sequence encoding a membrane domain as disclosed        herein (e.g., the wild-type sequence, functional fragment, or        variant thereof of the IL-12 polypeptide and/or membrane        domain), wherein the polynucleotide comprises at least one        chemically modified nucleobase, and wherein the polynucleotide        further comprises a miRNA binding site, e.g., a miRNA binding        site that binds to miR-122 (e.g., a miR-122-3p or miR-122-5p        binding site); and    -   (ii) a delivery agent comprising a compound having Formula (I).

In some embodiments, the polynucleotides, compositions or formulationsabove are used to treat and/or prevent an IL-12-related diseases,disorders or conditions, e.g., cancer.

28. Forms of Administration

The polynucleotides, pharmaceutical compositions and formulations of thedisclosure described above can be administered by any route that resultsin a therapeutically effective outcome. These include, but are notlimited to enteral (into the intestine), gastroenteral, epidural (intothe dura matter), oral (by way of the mouth), transdermal, peridural,intracerebral (into the cerebrum), intracerebroventricular (into thecerebral ventricles), epicutaneous (application onto the skin),intradermal, (into the skin itself), subcutaneous (under the skin),nasal administration (through the nose), intravenous (into a vein),intravenous bolus, intravenous drip, intraarterial (into an artery),intramuscular (into a muscle), intracardiac (into the heart),intraosseous infusion (into the bone marrow), intrathecal (into thespinal canal), intraperitoneal, (infusion or injection into theperitoneum), intravesical infusion, intravitreal, (through the eye),intracavernous injection (into a pathologic cavity) intracavitary (intothe base of the penis), intravaginal administration, intrauterine,extra-amniotic administration, transdermal (diffusion through the intactskin for systemic distribution), transmucosal (diffusion through amucous membrane), transvaginal, insufflation (snorting), sublingual,sublabial, enema, eye drops (onto the conjunctiva), in ear drops,auricular (in or by way of the ear), buccal (directed toward the cheek),conjunctival, cutaneous, dental (to a tooth or teeth), electro-osmosis,endocervical, endosinusial, endotracheal, extracorporeal, hemodialysis,infiltration, interstitial, intra-abdominal, intra-amniotic,intra-articular, intrabiliary, intrabronchial, intrabursal,intracartilaginous (within a cartilage), intracaudal (within the caudaequine), intracisternal (within the cisterna magna cerebellomedularis),intracorneal (within the cornea), dental intracornal, intracoronary(within the coronary arteries), intracorporus cavernosum (within thedilatable spaces of the corporus cavernosa of the penis), intradiscal(within a disc), intraductal (within a duct of a gland), intraduodenal(within the duodenum), intradural (within or beneath the dura),intraepidermal (to the epidermis), intraesophageal (to the esophagus),intragastric (within the stomach), intragingival (within the gingivae),intraileal (within the distal portion of the small intestine),intralesional (within or introduced directly to a localized lesion),intraluminal (within a lumen of a tube), intralymphatic (within thelymph), intramedullary (within the marrow cavity of a bone),intrameningeal (within the meninges), intraocular (within the eye),intraovarian (within the ovary), intrapericardial (within thepericardium), intrapleural (within the pleura), intraprostatic (withinthe prostate gland), intrapulmonary (within the lungs or its bronchi),intrasinal (within the nasal or periorbital sinuses), intraspinal(within the vertebral column), intrasynovial (within the synovial cavityof a joint), intratendinous (within a tendon), intratesticular (withinthe testicle), intrathecal (within the cerebrospinal fluid at any levelof the cerebrospinal axis), intrathoracic (within the thorax),intratubular (within the tubules of an organ), intratumor (within atumor), intratympanic (within the aurus media), intravascular (within avessel or vessels), intraventricular (within a ventricle), iontophoresis(by means of electric current where ions of soluble salts migrate intothe tissues of the body), irrigation (to bathe or flush open wounds orbody cavities), laryngeal (directly upon the larynx), nasogastric(through the nose and into the stomach), occlusive dressing technique(topical route administration that is then covered by a dressing thatoccludes the area), ophthalmic (to the external eye), oropharyngeal(directly to the mouth and pharynx), parenteral, percutaneous,periarticular, peridural, perineural, periodontal, rectal, respiratory(within the respiratory tract by inhaling orally or nasally for local orsystemic effect), retrobulbar (behind the pons or behind the eyeball),intramyocardial (entering the myocardium), soft tissue, subarachnoid,subconjunctival, submucosal, topical, transplacental (through or acrossthe placenta), transtracheal (through the wall of the trachea),transtympanic (across or through the tympanic cavity), ureteral (to theureter), urethral (to the urethra), vaginal, caudal block, diagnostic,nerve block, biliary perfusion, cardiac perfusion, photopheresis orspinal. In specific embodiments, compositions can be administered in away that allows them cross the blood-brain barrier, vascular barrier, orother epithelial barrier. In some embodiments, a formulation for a routeof administration can include at least one inactive ingredient.

The polynucleotides of the present disclosure can be delivered to a cellnaked. As used herein in, “naked” refers to delivering polynucleotidesfree from agents that promote transfection. For example, thepolynucleotides delivered to the cell may contain no modifications. Thenaked polynucleotides can be delivered to the cell using routes ofadministration known in the art and described herein.

The polynucleotides of the present disclosure can be formulated, usingthe methods described herein. The formulations can containpolynucleotides that can be modified and/or unmodified. The formulationsmay further include, but are not limited to, cell penetration agents, apharmaceutically acceptable carrier, a delivery agent, a bioerodible orbiocompatible polymer, a solvent, and a sustained-release deliverydepot. The formulated polynucleotides can be delivered to the cell usingroutes of administration known in the art and described herein.

The compositions can also be formulated for direct delivery to an organor tissue in any of several ways in the art including, but not limitedto, direct soaking or bathing, via a catheter, by gels, powder,ointments, creams, gels, lotions, and/or drops, by using substrates suchas fabric or biodegradable materials coated or impregnated with thecompositions, and the like.

The present disclosure encompasses the delivery of polynucleotides ofthe disclosure in forms suitable for parenteral and injectableadministration. Liquid dosage forms for parenteral administrationinclude, but are not limited to, pharmaceutically acceptable emulsions,microemulsions, solutions, suspensions, syrups, and/or elixirs. Inaddition to active ingredients, liquid dosage forms can comprise inertdiluents commonly used in the art such as, for example, water or othersolvents, solubilizing agents and emulsifiers such as ethyl alcohol,isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol,benzyl benzoate, propylene glycol, 1,3-butylene glycol,dimethylformamide, oils (in particular, cottonseed, groundnut, corn,germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfurylalcohol, polyethylene glycols and fatty acid esters of sorbitan, andmixtures thereof. Besides inert diluents, oral compositions can includeadjuvants such as wetting agents, emulsifying and suspending agents,sweetening, flavoring, and/or perfuming agents. In certain embodimentsfor parenteral administration, compositions are mixed with solubilizingagents such as CREMOPHOR®, alcohols, oils, modified oils, glycols,polysorbates, cyclodextrins, polymers, and/or combinations thereof.

A pharmaceutical composition for parenteral administration can compriseat least one inactive ingredient. Any or none of the inactiveingredients used may have been approved by the US Food and DrugAdministration (FDA). A non-exhaustive list of inactive ingredients foruse in pharmaceutical compositions for parenteral administrationincludes hydrochloric acid, mannitol, nitrogen, sodium acetate, sodiumchloride and sodium hydroxide.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions can be formulated according to the known artusing suitable dispersing agents, wetting agents, and/or suspendingagents. Sterile injectable preparations can be sterile injectablesolutions, suspensions, and/or emulsions in nontoxic parenterallyacceptable diluents and/or solvents, for example, as a solution in1,3-butanediol. Among the acceptable vehicles and solvents that can beemployed are water, Ringer's solution, U.S.P., and isotonic sodiumchloride solution. Sterile, fixed oils are conventionally employed as asolvent or suspending medium. For this purpose any bland fixed oil canbe employed including synthetic mono- or diglycerides. Fatty acids suchas oleic acid can be used in the preparation of injectables. The sterileformulation may also comprise adjuvants such as local anesthetics,preservatives and buffering agents.

Injectable formulations can be sterilized, for example, by filtrationthrough a bacterial-retaining filter, and/or by incorporatingsterilizing agents in the form of sterile solid compositions that can bedissolved or dispersed in sterile water or other sterile injectablemedium prior to use. Injectable formulations can be for direct injectioninto a region of a tissue, organ and/or subject. As a non-limitingexample, a tissue, organ and/or subject can be directly injected aformulation by intramyocardial injection into the ischemic region. (See,e.g., Zangi et al. Nature Biotechnology 2013; the contents of which areherein incorporated by reference in its entirety).

In order to prolong the effect of an active ingredient, it is oftendesirable to slow the absorption of the active ingredient fromsubcutaneous or intramuscular injection. This can be accomplished by theuse of a liquid suspension of crystalline or amorphous material withpoor water solubility. The rate of absorption of the drug then dependsupon its rate of dissolution which, in turn, may depend upon crystalsize and crystalline form. Alternatively, delayed absorption of aparenterally administered drug form is accomplished by dissolving orsuspending the drug in an oil vehicle. Injectable depot forms are madeby forming microencapsule matrices of the drug in biodegradable polymerssuch as polylactide-polyglycolide. Depending upon the ratio of drug topolymer and the nature of the particular polymer employed, the rate ofdrug release can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are prepared by entrapping the drug in liposomes ormicroemulsions that are compatible with body tissues.

29. Kits and Devices

a. Kits

The disclosure provides a variety of kits for conveniently and/oreffectively using the claimed nucleotides of the present disclosure.Typically kits will comprise sufficient amounts and/or numbers ofcomponents to allow a user to perform multiple treatments of asubject(s) and/or to perform multiple experiments.

In one aspect, the present disclosure provides kits comprising themolecules (polynucleotides) of the disclosure.

Said kits can be for protein production, comprising a firstpolynucleotides comprising a translatable region. The kit may furthercomprise packaging and instructions and/or a delivery agent to form aformulation composition. The delivery agent can comprise a saline, abuffered solution, a lipidoid or any delivery agent disclosed herein.

In some embodiments, the buffer solution can include sodium chloride,calcium chloride, phosphate and/or EDTA. In another embodiment, thebuffer solution can include, but is not limited to, saline, saline with2 mM calcium, 5% sucrose, 5% sucrose with 2 mM calcium, 5% Mannitol, 5%Mannitol with 2 mM calcium, Ringer's lactate, sodium chloride, sodiumchloride with 2 mM calcium and mannose (See, e.g., U.S. Pub. No.20120258046; herein incorporated by reference in its entirety). In afurther embodiment, the buffer solutions can be precipitated or it canbe lyophilized. The amount of each component can be varied to enableconsistent, reproducible higher concentration saline or simple bufferformulations. The components may also be varied in order to increase thestability of modified RNA in the buffer solution over a period of timeand/or under a variety of conditions. In one aspect, the presentdisclosure provides kits for protein production, comprising: apolynucleotide comprising a translatable region, provided in an amounteffective to produce a desired amount of a protein encoded by thetranslatable region when introduced into a target cell; a secondpolynucleotide comprising an inhibitory nucleic acid, provided in anamount effective to substantially inhibit the innate immune response ofthe cell; and packaging and instructions.

In one aspect, the present disclosure provides kits for proteinproduction, comprising a polynucleotide comprising a translatableregion, wherein the polynucleotide exhibits reduced degradation by acellular nuclease, and packaging and instructions.

In one aspect, the present disclosure provides kits for proteinproduction, comprising a polynucleotide comprising a translatableregion, wherein the polynucleotide exhibits reduced degradation by acellular nuclease, and a mammalian cell suitable for translation of thetranslatable region of the first nucleic acid.

b. Devices

The present disclosure provides for devices that may incorporatepolynucleotides that encode polypeptides of interest. These devicescontain in a stable formulation the reagents to synthesize apolynucleotide in a formulation available to be immediately delivered toa subject in need thereof, such as a human patient

Devices for administration can be employed to deliver thepolynucleotides of the present disclosure according to single, multi- orsplit-dosing regimens taught herein. Such devices are taught in, forexample, International Application Publ. No. WO2013151666, the contentsof which are incorporated herein by reference in their entirety.

Method and devices known in the art for multi-administration to cells,organs and tissues are contemplated for use in conjunction with themethods and compositions disclosed herein as embodiments of the presentdisclosure. These include, for example, those methods and devices havingmultiple needles, hybrid devices employing for example lumens orcatheters as well as devices utilizing heat, electric current orradiation driven mechanisms.

According to the present disclosure, these multi-administration devicescan be utilized to deliver the single, multi- or split dosescontemplated herein. Such devices are taught for example in,International Application Publ. No. WO2013151666, the contents of whichare incorporated herein by reference in their entirety.

In some embodiments, the polynucleotide is administered subcutaneouslyor intramuscularly via at least 3 needles to three different, optionallyadjacent, sites simultaneously, or within a 60 minutes period (e.g.,administration to 4, 5, 6, 7, 8, 9, or 10 sites simultaneously or withina 60 minute period).

c. Methods and Devices Utilizing Catheters and/or Lumens

Methods and devices using catheters and lumens can be employed toadminister the polynucleotides of the present disclosure on a single,multi- or split dosing schedule. Such methods and devices are describedin International Application Publication No. WO2013151666, the contentsof which are incorporated herein by reference in their entirety.

d. Methods and Devices Utilizing Electrical Current

Methods and devices utilizing electric current can be employed todeliver the polynucleotides of the present disclosure according to thesingle, multi- or split dosing regimens taught herein. Such methods anddevices are described in International Application Publication No.WO2013151666, the contents of which are incorporated herein by referencein their entirety.

30. Definitions

In order that the present disclosure can be more readily understood,certain terms are first defined. As used in this application, except asotherwise expressly provided herein, each of the following terms shallhave the meaning set forth below. Additional definitions are set forththroughout the application.

The disclosure includes embodiments in which exactly one member of thegroup is present in, employed in, or otherwise relevant to a givenproduct or process. The disclosure includes embodiments in which morethan one, or all of the group members are present in, employed in, orotherwise relevant to a given product or process.

In this specification and the appended claims, the singular forms “a”,“an” and “the” include plural referents unless the context clearlydictates otherwise. The terms “a” (or “an”), as well as the terms “oneor more,” and “at least one” can be used interchangeably herein. Incertain aspects, the term “a” or “an” means “single.” In other aspects,the term “a” or “an” includes “two or more” or “multiple.”

Furthermore, “and/or” where used herein is to be taken as specificdisclosure of each of the two specified features or components with orwithout the other. Thus, the term “and/or” as used in a phrase such as“A and/or B” herein is intended to include “A and B,” “A or B,” “A”(alone), and “B” (alone). Likewise, the term “and/or” as used in aphrase such as “A, B, and/or C” is intended to encompass each of thefollowing aspects: A, B, and C; A, B, or C; A or C; A or B; B or C; Aand C; A and B; B and C; A (alone); B (alone); and C (alone).

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this disclosure is related. For example, the ConciseDictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd ed.,2002, CRC Press; The Dictionary of Cell and Molecular Biology, 3rd ed.,1999, Academic Press; and the Oxford Dictionary Of Biochemistry AndMolecular Biology, Revised, 2000, Oxford University Press, provide oneof skill with a general dictionary of many of the terms used in thisdisclosure.

Wherever aspects are described herein with the language “comprising,”otherwise analogous aspects described in terms of “consisting of” and/or“consisting essentially of” are also provided. Units, prefixes, andsymbols are denoted in their Systeme International de Unites (SI)accepted form. Numeric ranges are inclusive of the numbers defining therange. Where a range of values is recited, it is to be understood thateach intervening integer value, and each fraction thereof, between therecited upper and lower limits of that range is also specificallydisclosed, along with each subrange between such values. The upper andlower limits of any range can independently be included in or excludedfrom the range, and each range where either, neither or both limits areincluded is also encompassed within the disclosure. Where a value isexplicitly recited, it is to be understood that values which are aboutthe same quantity or amount as the recited value are also within thescope of the disclosure. Where a combination is disclosed, eachsubcombination of the elements of that combination is also specificallydisclosed and is within the scope of the disclosure. Conversely, wheredifferent elements or groups of elements are individually disclosed,combinations thereof are also disclosed. Where any element of adisclosure is disclosed as having a plurality of alternatives, examplesof that disclosure in which each alternative is excluded singly or inany combination with the other alternatives are also hereby disclosed;more than one element of a disclosure can have such exclusions, and allcombinations of elements having such exclusions are hereby disclosed.

Nucleotides are referred to by their commonly accepted single-lettercodes. Unless otherwise indicated, nucleic acids are written left toright in 5′ to 3′ orientation. Nucleobases are referred to herein bytheir commonly known one-letter symbols recommended by the IUPAC-IUBBiochemical Nomenclature Commission. Accordingly, A represents adenine,C represents cytosine, G represents guanine, T represents thymine, Urepresents uracil.

Amino acids are referred to herein by either their commonly known threeletter symbols or by the one-letter symbols recommended by the IUPAC-IUBBiochemical Nomenclature Commission. Unless otherwise indicated, aminoacid sequences are written left to right in amino to carboxyorientation.

About: The term “about” as used in connection with a numerical valuethroughout the specification and the claims denotes an interval ofaccuracy, familiar and acceptable to a person skilled in the art. Ingeneral, such interval of accuracy is ±10%.

Where ranges are given, endpoints are included. Furthermore, unlessotherwise indicated or otherwise evident from the context andunderstanding of one of ordinary skill in the art, values that areexpressed as ranges can assume any specific value or subrange within thestated ranges in different embodiments of the disclosure, to the tenthof the unit of the lower limit of the range, unless the context clearlydictates otherwise.

Administered in combination: As used herein, the term “administered incombination” or “combined administration” means that two or more agentsare administered to a subject at the same time or within an intervalsuch that there can be an overlap of an effect of each agent on thepatient. In some embodiments, they are administered within about 60, 30,15, 10, 5, or 1 minute of one another. In some embodiments, theadministrations of the agents are spaced sufficiently closely togethersuch that a combinatorial (e.g., a synergistic) effect is achieved.

Amino acid substitution: The term “amino acid substitution” refers toreplacing an amino acid residue present in a parent or referencesequence (e.g., a wild type IL-12 sequence) with another amino acidresidue. An amino acid can be substituted in a parent or referencesequence (e.g., a wild type IL-12 polypeptide sequence), for example,via chemical peptide synthesis or through recombinant methods known inthe art. Accordingly, a reference to a “substitution at position X”refers to the substitution of an amino acid present at position X withan alternative amino acid residue. In some aspects, substitutionpatterns can be described according to the schema AnY, wherein A is thesingle letter code corresponding to the amino acid naturally ororiginally present at position n, and Y is the substituting amino acidresidue. In other aspects, substitution patterns can be describedaccording to the schema An(YZ), wherein A is the single letter codecorresponding to the amino acid residue substituting the amino acidnaturally or originally present at position X, and Y and Z arealternative substituting amino acid residue, i.e.,

In the context of the present disclosure, substitutions (even when theyreferred to as amino acid substitution) are conducted at the nucleicacid level, i.e., substituting an amino acid residue with an alternativeamino acid residue is conducted by substituting the codon encoding thefirst amino acid with a codon encoding the second amino acid.

Animal: As used herein, the term “animal” refers to any member of theanimal kingdom. In some embodiments, “animal” refers to humans at anystage of development. In some embodiments, “animal” refers to non-humananimals at any stage of development. In certain embodiments, thenon-human animal is a mammal (e.g., a rodent, a mouse, a rat, a rabbit,a monkey, a dog, a cat, a sheep, cattle, a primate, or a pig). In someembodiments, animals include, but are not limited to, mammals, birds,reptiles, amphibians, fish, and worms. In some embodiments, the animalis a transgenic animal, genetically-engineered animal, or a clone.

Approximately: As used herein, the term “approximately,” as applied toone or more values of interest, refers to a value that is similar to astated reference value. In certain embodiments, the term “approximately”refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%,16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%,or less in either direction (greater than or less than) of the statedreference value unless otherwise stated or otherwise evident from thecontext (except where such number would exceed 100% of a possiblevalue).

Associated with: As used herein with respect to a disease, the term“associated with” means that the symptom, measurement, characteristic,or status in question is linked to the diagnosis, development, presence,or progression of that disease. As association may, but need not, becausatively linked to the disease. For example, symptoms, sequelae, orany effects causing a decrease in the quality of life of a patient ofcancer are considered associated with cancer and in some embodiments ofthe present disclosure can be treated, ameliorated, or prevented byadministering the polynucleotides of the present disclosure to a subjectin need thereof.

When used with respect to two or more moieties, the terms “associatedwith,” “conjugated,” “linked,” “attached,” and “tethered,” when usedwith respect to two or more moieties, means that the moieties arephysically associated or connected with one another, either directly orvia one or more additional moieties that serves as a linking agent, toform a structure that is sufficiently stable so that the moieties remainphysically associated under the conditions in which the structure isused, e.g., physiological conditions. An “association” need not bestrictly through direct covalent chemical bonding. It may also suggestionic or hydrogen bonding or a hybridization based connectivitysufficiently stable such that the “associated” entities remainphysically associated.

Bifunctional: As used herein, the term “bifunctional” refers to anysubstance, molecule or moiety that is capable of or maintains at leasttwo functions. The functions may affect the same outcome or a differentoutcome. The structure that produces the function can be the same ordifferent. For example, bifunctional modified RNAs of the presentdisclosure may encode an IL-12 peptide (a first function) while thosenucleosides that comprise the encoding RNA are, in and of themselves,capable of extending the half-life of the RNA (second function). In thisexample, delivery of the bifunctional modified RNA to a subjectsuffering from a protein deficiency would produce not only a peptide orprotein molecule that may ameliorate or treat a disease or conditions,but would also maintain a population modified RNA present in the subjectfor a prolonged period of time. In other aspects, a bifunctionallymodified mRNA can be a chimeric molecule comprising, for example, an RNAencoding an IL-12 peptide (a first function) and a second protein eitherfused to first protein or co-expressed with the first protein.

Biocompatible: As used herein, the term “biocompatible” means compatiblewith living cells, tissues, organs or systems posing little to no riskof injury, toxicity or rejection by the immune system.

Biodegradable: As used herein, the term “biodegradable” means capable ofbeing broken down into innocuous products by the action of livingthings.

Biologically active: As used herein, the phrase “biologically active”refers to a characteristic of any substance that has activity in abiological system and/or organism. For instance, a substance that, whenadministered to an organism, has a biological effect on that organism,is considered to be biologically active. In particular embodiments, apolynucleotide of the present disclosure can be considered biologicallyactive if even a portion of the polynucleotide is biologically active ormimics an activity considered biologically relevant.

Chimera: As used herein, “chimera” is an entity having two or moreincongruous or heterogeneous parts or regions. For example, a chimericmolecule can comprise a first part comprising an IL-12B polypeptide,IL-12A polypeptide, or both IL-12B and IL-12A polypeptides, and a secondpart (e.g., genetically fused or linked to the first part) comprising amembrane domain. A chimera can also include a tethered IL-12 polypeptideas disclosed herein further comprising a second therapeutic protein(e.g., a protein with a distinct enzymatic activity, an antigen bindingmoiety, or a moiety capable of extending the plasma half-life of IL-12,for example, an Fc region of an antibody).

Sequence Optimization: The term “sequence optimization” refers to aprocess or series of processes by which nucleobases in a referencenucleic acid sequence are replaced with alternative nucleobases,resulting in a nucleic acid sequence with improved properties, e.g.,improved protein expression or decreased immunogenicity.

In general, the goal in sequence optimization is to produce a synonymousnucleotide sequence than encodes the same polypeptide sequence encodedby the reference nucleotide sequence. Thus, there are no amino acidsubstitutions (as a result of codon optimization) in the polypeptideencoded by the codon optimized nucleotide sequence with respect to thepolypeptide encoded by the reference nucleotide sequence.

Codon substitution: The terms “codon substitution” or “codonreplacement” in the context of sequence optimization refer to replacinga codon present in a reference nucleic acid sequence with another codon.A codon can be substituted in a reference nucleic acid sequence, forexample, via chemical peptide synthesis or through recombinant methodsknown in the art. Accordingly, references to a “substitution” or“replacement” at a certain location in a nucleic acid sequence (e.g., anmRNA) or within a certain region or subsequence of a nucleic acidsequence (e.g., an mRNA) refer to the substitution of a codon at suchlocation or region with an alternative codon.

As used herein, the terms “coding region” and “region encoding” andgrammatical variants thereof, refer to an Open Reading Frame (ORF) in apolynucleotide that upon expression yields a polypeptide or protein.

Compound: As used herein, the term “compound,” is meant to include allstereoisomers and isotopes of the structure depicted. As used herein,the term “stereoisomer” means any geometric isomer (e.g., cis- andtrans-isomer), enantiomer, or diastereomer of a compound. The presentdisclosure encompasses any and all stereoisomers of the compoundsdescribed herein, including stereomerically pure forms (e.g.,geometrically pure, enantiomerically pure, or diastereomerically pure)and enantiomeric and stereoisomeric mixtures, e.g., racemates.Enantiomeric and stereomeric mixtures of compounds and means ofresolving them into their component enantiomers or stereoisomers arewell-known. “Isotopes” refers to atoms having the same atomic number butdifferent mass numbers resulting from a different number of neutrons inthe nuclei. For example, isotopes of hydrogen include tritium anddeuterium. Further, a compound, salt, or complex of the presentdisclosure can be prepared in combination with solvent or watermolecules to form solvates and hydrates by routine methods.

Contacting: As used herein, the term “contacting” means establishing aphysical connection between two or more entities. For example,contacting a mammalian cell with a nanoparticle composition means thatthe mammalian cell and a nanoparticle are made to share a physicalconnection. Methods of contacting cells with external entities both invivo and ex vivo are well known in the biological arts. For example,contacting a nanoparticle composition and a mammalian cell disposedwithin a mammal may be performed by varied routes of administration(e.g., intravenous, intramuscular, intradermal, and subcutaneous) andmay involve varied amounts of nanoparticle compositions. Moreover, morethan one mammalian cell may be contacted by a nanoparticle composition.

Conservative amino acid substitution: A “conservative amino acidsubstitution” is one in which the amino acid residue in a proteinsequence is replaced with an amino acid residue having a similar sidechain. Families of amino acid residues having similar side chains havebeen defined in the art, including basic side chains (e.g., lysine,arginine, or histidine), acidic side chains (e.g., aspartic acid orglutamic acid), uncharged polar side chains (e.g., glycine, asparagine,glutamine, serine, threonine, tyrosine, or cysteine), nonpolar sidechains (e.g., alanine, valine, leucine, isoleucine, proline,phenylalanine, methionine, or tryptophan), beta-branched side chains(e.g., threonine, valine, isoleucine) and aromatic side chains (e.g.,tyrosine, phenylalanine, tryptophan, or histidine). Thus, if an aminoacid in a polypeptide is replaced with another amino acid from the sameside chain family, the amino acid substitution is considered to beconservative. In another aspect, a string of amino acids can beconservatively replaced with a structurally similar string that differsin order and/or composition of side chain family members.

Non-conservative amino acid substitution: Non-conservative amino acidsubstitutions include those in which (i) a residue having anelectropositive side chain (e.g., Arg, His or Lys) is substituted for,or by, an electronegative residue (e.g., Glu or Asp), (ii) a hydrophilicresidue (e.g., Ser or Thr) is substituted for, or by, a hydrophobicresidue (e.g., Ala, Leu, Ile, Phe or Val), (iii) a cysteine or prolineis substituted for, or by, any other residue, or (iv) a residue having abulky hydrophobic or aromatic side chain (e.g., Val, His, Ile or Trp) issubstituted for, or by, one having a smaller side chain (e.g., Ala orSer) or no side chain (e.g., Gly).

Other amino acid substitutions can be readily identified by workers ofordinary skill. For example, for the amino acid alanine, a substitutioncan be taken from any one of D-alanine, glycine, beta-alanine,L-cysteine and D-cysteine. For lysine, a replacement can be any one ofD-lysine, arginine, D-arginine, homo-arginine, methionine, D-methionine,ornithine, or D-ornithine. Generally, substitutions in functionallyimportant regions that can be expected to induce changes in theproperties of isolated polypeptides are those in which (i) a polarresidue, e.g., serine or threonine, is substituted for (or by) ahydrophobic residue, e.g., leucine, isoleucine, phenylalanine, oralanine; (ii) a cysteine residue is substituted for (or by) any otherresidue; (iii) a residue having an electropositive side chain, e.g.,lysine, arginine or histidine, is substituted for (or by) a residuehaving an electronegative side chain, e.g., glutamic acid or asparticacid; or (iv) a residue having a bulky side chain, e.g., phenylalanine,is substituted for (or by) one not having such a side chain, e.g.,glycine. The likelihood that one of the foregoing non-conservativesubstitutions can alter functional properties of the protein is alsocorrelated to the position of the substitution with respect tofunctionally important regions of the protein: some non-conservativesubstitutions can accordingly have little or no effect on biologicalproperties.

Conserved: As used herein, the term “conserved” refers to nucleotides oramino acid residues of a polynucleotide sequence or polypeptidesequence, respectively, that are those that occur unaltered in the sameposition of two or more sequences being compared. Nucleotides or aminoacids that are relatively conserved are those that are conserved amongstmore related sequences than nucleotides or amino acids appearingelsewhere in the sequences.

In some embodiments, two or more sequences are said to be “completelyconserved” if they are 100% identical to one another. In someembodiments, two or more sequences are said to be “highly conserved” ifthey are at least 70% identical, at least 80% identical, at least 90%identical, or at least 95% identical to one another. In someembodiments, two or more sequences are said to be “highly conserved” ifthey are about 70% identical, about 80% identical, about 90% identical,about 95%, about 98%, or about 99% identical to one another. In someembodiments, two or more sequences are said to be “conserved” if theyare at least 30% identical, at least 40% identical, at least 50%identical, at least 60% identical, at least 70% identical, at least 80%identical, at least 90% identical, or at least 95% identical to oneanother. In some embodiments, two or more sequences are said to be“conserved” if they are about 30% identical, about 40% identical, about50% identical, about 60% identical, about 70% identical, about 80%identical, about 90% identical, about 95% identical, about 98%identical, or about 99% identical to one another. Conservation ofsequence may apply to the entire length of a polynucleotide orpolypeptide or may apply to a portion, region or feature thereof.

Controlled Release: As used herein, the term “controlled release” refersto a pharmaceutical composition or compound release profile thatconforms to a particular pattern of release to effect a therapeuticoutcome.

Cyclic or Cyclized: As used herein, the term “cyclic” refers to thepresence of a continuous loop. Cyclic molecules need not be circular,only joined to form an unbroken chain of subunits. Cyclic molecules suchas the engineered RNA or mRNA of the present disclosure can be singleunits or multimers or comprise one or more components of a complex orhigher order structure.

Cytotoxic: As used herein, “cytotoxic” refers to killing or causinginjurious, toxic, or deadly effect on a cell (e.g., a mammalian cell(e.g., a human cell)), bacterium, virus, fungus, protozoan, parasite,prion, or a combination thereof.

Delivering: As used herein, the term “delivering” means providing anentity to a destination. For example, delivering a polynucleotide to asubject may involve administering a nanoparticle composition includingthe polynucleotide to the subject (e.g., by an intravenous,intramuscular, intradermal, or subcutaneous route). Administration of ananoparticle composition to a mammal or mammalian cell may involvecontacting one or more cells with the nanoparticle composition.

Delivery Agent: As used herein, “delivery agent” refers to any substancethat facilitates, at least in part, the in vivo, in vitro, or ex vivodelivery of a polynucleotide to targeted cells.

Destabilized: As used herein, the term “destable,” “destabilize,” or“destabilizing region” means a region or molecule that is less stablethan a starting, wild-type or native form of the same region ormolecule.

Diastereomer: As used herein, the term “diastereomer,” meansstereoisomers that are not mirror images of one another and arenon-superimposable on one another.

Digest: As used herein, the term “digest” means to break apart intosmaller pieces or components. When referring to polypeptides orproteins, digestion results in the production of peptides.

Distal: As used herein, the term “distal” means situated away from thecenter or away from a point or region of interest.

Domain: As used herein, when referring to polypeptides, the term“domain” refers to a motif of a polypeptide having one or moreidentifiable structural or functional characteristics or properties(e.g., binding capacity, serving as a site for protein-proteininteractions).

Dosing regimen: As used herein, a “dosing regimen” or a “dosing regimen”is a schedule of administration or physician determined regimen oftreatment, prophylaxis, or palliative care.

Effective Amount: As used herein, the term “effective amount” of anagent is that amount sufficient to effect beneficial or desired results,for example, clinical results, and, as such, an “effective amount”depends upon the context in which it is being applied. For example, inthe context of administering an agent that treats a protein deficiency(e.g., an IL-12 deficiency), an effective amount of an agent is, forexample, an amount of mRNA expressing sufficient PBGF to ameliorate,reduce, eliminate, or prevent the symptoms associated with the IL-12deficiency, as compared to the severity of the symptom observed withoutadministration of the agent. The term “effective amount” can be usedinterchangeably with “effective dose,” “therapeutically effectiveamount,” or “therapeutically effective dose.”

Enantiomer: As used herein, the term “enantiomer” means each individualoptically active form of a compound of the disclosure, having an opticalpurity or enantiomeric excess (as determined by methods standard in theart) of at least 80% (i.e., at least 90% of one enantiomer and at most10% of the other enantiomer), at least 90%, or at least 98%.

Encapsulate: As used herein, the term “encapsulate” means to enclose,surround or encase.

Encapsulation Efficiency: As used herein, “encapsulation efficiency”refers to the amount of a polynucleotide that becomes part of ananoparticle composition, relative to the initial total amount ofpolynucleotide used in the preparation of a nanoparticle composition.For example, if 97 mg of polynucleotide are encapsulated in ananoparticle composition out of a total 100 mg of polynucleotideinitially provided to the composition, the encapsulation efficiency maybe given as 97%. As used herein, “encapsulation” may refer to complete,substantial, or partial enclosure, confinement, surrounding, orencasement.

Encoded protein cleavage signal: As used herein, “encoded proteincleavage signal” refers to the nucleotide sequence that encodes aprotein cleavage signal.

Engineered: As used herein, embodiments of the disclosure are“engineered” when they are designed to have a feature or property,whether structural or chemical, that varies from a starting point, wildtype or native molecule.

Enhanced Delivery: As used herein, the term “enhanced delivery” meansdelivery of more (e.g., at least 1.5 fold more, at least 2-fold more, atleast 3-fold more, at least 4-fold more, at least 5-fold more, at least6-fold more, at least 7-fold more, at least 8-fold more, at least 9-foldmore, at least 10-fold more) of a polynucleotide by a nanoparticle to atarget tissue of interest (e.g., mammalian liver) compared to the levelof delivery of a polynucleotide by a control nanoparticle to a targettissue of interest (e.g., MC3, KC2, or DLinDMA). The level of deliveryof a nanoparticle to a particular tissue may be measured by comparingthe amount of protein produced in a tissue to the weight of said tissue,comparing the amount of polynucleotide in a tissue to the weight of saidtissue, comparing the amount of protein produced in a tissue to theamount of total protein in said tissue, or comparing the amount ofpolynucleotide in a tissue to the amount of total polynucleotide in saidtissue. It will be understood that the enhanced delivery of ananoparticle to a target tissue need not be determined in a subjectbeing treated, it may be determined in a surrogate such as an animalmodel (e.g., a rat model).

Exosome: As used herein, “exosome” is a vesicle secreted by mammaliancells or a complex involved in RNA degradation.

Expression: As used herein, “expression” of a nucleic acid sequencerefers to one or more of the following events: (1) production of an mRNAtemplate from a DNA sequence (e.g., by transcription); (2) processing ofan mRNA transcript (e.g., by splicing, editing, 5′ cap formation, and/or3′ end processing); (3) translation of an mRNA into a polypeptide orprotein; and (4) post-translational modification of a polypeptide orprotein.

Ex Vivo: As used herein, the term “ex vivo” refers to events that occuroutside of an organism (e.g., animal, plant, or microbe or cell ortissue thereof). Ex vivo events may take place in an environmentminimally altered from a natural (e.g., in vivo) environment.

Feature: As used herein, a “feature” refers to a characteristic, aproperty, or a distinctive element. When referring to polypeptides,“features” are defined as distinct amino acid sequence-based componentsof a molecule. Features of the polypeptides encoded by thepolynucleotides of the present disclosure include surfacemanifestations, local conformational shape, folds, loops, half-loops,domains, half-domains, sites, termini or any combination thereof.

Formulation: As used herein, a “formulation” includes at least apolynucleotide and one or more of a carrier, an excipient, and adelivery agent.

Fragment: A “fragment,” as used herein, refers to a portion. Forexample, fragments of proteins can comprise polypeptides obtained bydigesting full-length protein isolated from cultured cells. In someembodiments, a fragment is a subsequence of a full length protein (e.g.,IL-12 and/or a membrane domain) wherein N-terminal, and/or C-terminal,and/or internal subsequences have been deleted. In some preferredaspects of the present disclosure, the fragments of a protein of thepresent disclosure are functional fragments.

Functional: As used herein, a “functional” biological molecule is abiological molecule in a form in which it exhibits a property and/oractivity by which it is characterized. Thus, a functional fragment of apolynucleotide of the present disclosure is a polynucleotide capable ofexpressing a functional IL-12 and/or membrane domain fragment. As usedherein, a functional fragment of IL-12 refers to a fragment of wild typeIL-12 (i.e., a fragment of any of its naturally occurring isoforms), ora mutant or variant thereof, wherein the fragment retains a least about10%, at least about 15%, at least about 20%, at least about 25%, atleast about 30%, at least about 35%, at least about 40%, at least about45%, at least about 50%, at least about 55%, at least about 60%, atleast about 65%, at least about 70%, at least about 75%, at least about80%, at least about 85%, at least about 90%, or at least about 95% ofthe biological activity of the corresponding full length protein. Asused herein, a functional fragment of a membrane domain is a fragment ofa wild type membrane domain, or a mutant or variant thereof, wherein thefragment is capable of tethering an IL-12 polypeptide to a cellmembrane.

Helper Lipid: As used herein, the term “helper lipid” refers to acompound or molecule that includes a lipidic moiety (for insertion intoa lipid layer, e.g., lipid bilayer) and a polar moiety (for interactionwith physiologic solution at the surface of the lipid layer). Typicallythe helper lipid is a phospholipid. A function of the helper lipid is to“complement” the amino lipid and increase the fusogenicity of thebilayer and/or to help facilitate endosomal escape, e.g., of nucleicacid delivered to cells. Helper lipids are also believed to be a keystructural component to the surface of the LNP.

Homology: As used herein, the term “homology” refers to the overallrelatedness between polymeric molecules, e.g. between nucleic acidmolecules (e.g. DNA molecules and/or RNA molecules) and/or betweenpolypeptide molecules. Generally, the term “homology” implies anevolutionary relationship between two molecules. Thus, two moleculesthat are homologous will have a common evolutionary ancestor. In thecontext of the present disclosure, the term homology encompasses both toidentity and similarity.

In some embodiments, polymeric molecules are considered to be“homologous” to one another if at least 25%, 30%, 35%, 40%, 45%, 50%,55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% of the monomers inthe molecule are identical (exactly the same monomer) or are similar(conservative substitutions). The term “homologous” necessarily refersto a comparison between at least two sequences (polynucleotide orpolypeptide sequences).

Identity: As used herein, the term “identity” refers to the overallmonomer conservation between polymeric molecules, e.g., betweenpolynucleotide molecules (e.g. DNA molecules and/or RNA molecules)and/or between polypeptide molecules. Calculation of the percentidentity of two polynucleotide sequences, for example, can be performedby aligning the two sequences for optimal comparison purposes (e.g.,gaps can be introduced in one or both of a first and a second nucleicacid sequences for optimal alignment and non-identical sequences can bedisregarded for comparison purposes). In certain embodiments, the lengthof a sequence aligned for comparison purposes is at least 30%, at least40%, at least 50%, at least 60%, at least 70%, at least 80%, at least90%, at least 95%, or 100% of the length of the reference sequence. Thenucleotides at corresponding nucleotide positions are then compared.When a position in the first sequence is occupied by the same nucleotideas the corresponding position in the second sequence, then the moleculesare identical at that position. The percent identity between the twosequences is a function of the number of identical positions shared bythe sequences, taking into account the number of gaps, and the length ofeach gap, which needs to be introduced for optimal alignment of the twosequences. The comparison of sequences and determination of percentidentity between two sequences can be accomplished using a mathematicalalgorithm. When comparing DNA and RNA, thymine (T) and uracil (U) can beconsidered equivalent.

Suitable software programs are available from various sources, and foralignment of both protein and nucleotide sequences. One suitable programto determine percent sequence identity is bl2seq, part of the BLASTsuite of program available from the U.S. government's National Centerfor Biotechnology Information BLAST web site (blast.ncbi.nlm.nih.gov).Bl2seq performs a comparison between two sequences using either theBLASTN or BLASTP algorithm. BLASTN is used to compare nucleic acidsequences, while BLASTP is used to compare amino acid sequences. Othersuitable programs are, e.g., Needle, Stretcher, Water, or Matcher, partof the EMBOSS suite of bioinformatics programs and also available fromthe European Bioinformatics Institute (EBI) at www.ebi.ac.uk/Tools/psa.

Sequence alignments can be conducted using methods known in the art suchas MAFFT, Clustal (ClustalW, Clustal X or Clustal Omega), MUSCLE, etc.

Different regions within a single polynucleotide or polypeptide targetsequence that aligns with a polynucleotide or polypeptide referencesequence can each have their own percent sequence identity. It is notedthat the percent sequence identity value is rounded to the nearesttenth. For example, 80.11, 80.12, 80.13, and 80.14 are rounded down to80.1, while 80.15, 80.16, 80.17, 80.18, and 80.19 are rounded up to80.2. It also is noted that the length value will always be an integer.

In certain aspects, the percentage identity “% ID” of a first amino acidsequence (or nucleic acid sequence) to a second amino acid sequence (ornucleic acid sequence) is calculated as % ID=100×(Y/Z), where Y is thenumber of amino acid residues (or nucleobases) scored as identicalmatches in the alignment of the first and second sequences (as alignedby visual inspection or a particular sequence alignment program) and Zis the total number of residues in the second sequence. If the length ofa first sequence is longer than the second sequence, the percentidentity of the first sequence to the second sequence will be higherthan the percent identity of the second sequence to the first sequence.

One skilled in the art will appreciate that the generation of a sequencealignment for the calculation of a percent sequence identity is notlimited to binary sequence-sequence comparisons exclusively driven byprimary sequence data. It will also be appreciated that sequencealignments can be generated by integrating sequence data with data fromheterogeneous sources such as structural data (e.g., crystallographicprotein structures), functional data (e.g., location of mutations), orphylogenetic data. A suitable program that integrates heterogeneous datato generate a multiple sequence alignment is T-Coffee, available atwww.tcoffee.org, and alternatively available, e.g., from the EBI. Itwill also be appreciated that the final alignment used to calculatepercent sequence identity can be curated either automatically ormanually.

Immune response: The term “immune response” refers to the action of, forexample, lymphocytes, antigen presenting cells, phagocytic cells,granulocytes, and soluble macromolecules produced by the above cells orthe liver (including antibodies, cytokines, and complement) that resultsin selective damage to, destruction of, or elimination from the humanbody of invading pathogens, cells or tissues infected with pathogens,cancerous cells, or, in cases of autoimmunity or pathologicalinflammation, normal human cells or tissues. In some cases, theadministration of a nanoparticle comprising a lipid component and anencapsulated therapeutic agent can trigger an immune response, which canbe caused by (i) the encapsulated therapeutic agent (e.g., an mRNA),(ii) the expression product of such encapsulated therapeutic agent(e.g., a polypeptide encoded by the mRNA), (iii) the lipid component ofthe nanoparticle, or (iv) a combination thereof. Inflammatory response:“Inflammatory response” refers to immune responses involving specificand non-specific defense systems. A specific defense system reaction isa specific immune system reaction to an antigen. Examples of specificdefense system reactions include antibody responses. A non-specificdefense system reaction is an inflammatory response mediated byleukocytes generally incapable of immunological memory, e.g.,macrophages, eosinophils and neutrophils. In some aspects, an immuneresponse includes the secretion of inflammatory cytokines, resulting inelevated inflammatory cytokine levels.

Inflammatory cytokines: The term “inflammatory cytokine” refers tocytokines that are elevated in an inflammatory response. Examples ofinflammatory cytokines include interleukin-6 (IL-6), CXCL1 (chemokine(C-X-C motif) ligand 1; also known as GROα, interferon-γ (IFNγ), tumornecrosis factor α (TNFα), interferon γ-induced protein 10 (IP-10), orgranulocyte-colony stimulating factor (G-CSF). The term inflammatorycytokines includes also other cytokines associated with inflammatoryresponses known in the art, e.g., interleukin-1 (IL-1), interleukin-8(IL-8), interleukin-12 (IL-12), interleukin-13 (I1-13), interferon α(IFN-α), etc.

In Vitro: As used herein, the term “in vitro” refers to events thatoccur in an artificial environment, e.g., in a test tube or reactionvessel, in cell culture, in a Petri dish, etc., rather than within anorganism (e.g., animal, plant, or microbe).

In Vivo: As used herein, the term “in vivo” refers to events that occurwithin an organism (e.g., animal, plant, or microbe or cell or tissuethereof).

Insertional and deletional variants: “Insertional variants” whenreferring to polypeptides are those with one or more amino acidsinserted immediately adjacent to an amino acid at a particular positionin a native or starting sequence. “Immediately adjacent” to an aminoacid means connected to either the alpha-carboxy or alpha-aminofunctional group of the amino acid. “Deletional variants” when referringto polypeptides are those with one or more amino acids in the native orstarting amino acid sequence removed. Ordinarily, deletional variantswill have one or more amino acids deleted in a particular region of themolecule.

Intact: As used herein, in the context of a polypeptide, the term“intact” means retaining an amino acid corresponding to the wild typeprotein, e.g., not mutating or substituting the wild type amino acid.Conversely, in the context of a nucleic acid, the term “intact” meansretaining a nucleobase corresponding to the wild type nucleic acid,e.g., not mutating or substituting the wild type nucleobase.

Intracellular domain: As used herein, the terms “intracellular domain”,“IC” and “ICD” refer to the region of a polypeptide located inside acell. In some embodiments, an intracellular domain transmits a signal tothe cell. In some embodiments, the tethered IL-12 polypeptides encodedby the polynucleotides (e.g., mRNA) described herein, comprise anintracellular domain that transmits a signal to the cell. In someembodiments, the tethered IL-12 polypeptides encoded by thepolynucleotides (e.g., mRNA) described herein, comprise an intracellulardomain that does not transmit a signal to the cell.

Ionizable amino lipid: The term “ionizable amino lipid” includes thoselipids having one, two, three, or more fatty acid or fatty alkyl chainsand a pH-titratable amino head group (e.g., an alkylamino ordialkylamino head group). An ionizable amino lipid is typicallyprotonated (i.e., positively charged) at a pH below the pKa of the aminohead group and is substantially not charged at a pH above the pKa. Suchionizable amino lipids include, but are not limited to DLin-MC3-DMA(MC3) and (13Z,165Z)-N,N-dimethyl-3-nonydocosa-13-16-dien-1-amine(L608).

Isolated: As used herein, the term “isolated” refers to a substance orentity that has been separated from at least some of the components withwhich it was associated (whether in nature or in an experimentalsetting). Isolated substances (e.g., polynucleotides or polypeptides)can have varying levels of purity in reference to the substances fromwhich they have been isolated. Isolated substances and/or entities canbe separated from at least about 10%, about 20%, about 30%, about 40%,about 50%, about 60%, about 70%, about 80%, about 90%, or more of theother components with which they were initially associated. In someembodiments, isolated substances are more than about 80%, about 85%,about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about96%, about 97%, about 98%, about 99%, or more than about 99% pure. Asused herein, a substance is “pure” if it is substantially free of othercomponents.

Substantially isolated: By “substantially isolated” is meant that thecompound is substantially separated from the environment in which it wasformed or detected. Partial separation can include, for example, acomposition enriched in the compound of the present disclosure.Substantial separation can include compositions containing at leastabout 50%, at least about 60%, at least about 70%, at least about 80%,at least about 90%, at least about 95%, at least about 97%, or at leastabout 99% by weight of the compound of the present disclosure, or saltthereof.

A polynucleotide, vector, polypeptide, cell, or any compositiondisclosed herein which is “isolated” is a polynucleotide, vector,polypeptide, cell, or composition which is in a form not found innature. Isolated polynucleotides, vectors, polypeptides, or compositionsinclude those which have been purified to a degree that they are nolonger in a form in which they are found in nature. In some aspects, apolynucleotide, vector, polypeptide, or composition which is isolated issubstantially pure.

Isomer: As used herein, the term “isomer” means any tautomer,stereoisomer, enantiomer, or diastereomer of any compound of thedisclosure. It is recognized that the compounds of the disclosure canhave one or more chiral centers and/or double bonds and, therefore,exist as stereoisomers, such as double-bond isomers (i.e., geometric E/Zisomers) or diastereomers (e.g., enantiomers (i.e., (+) or (−)) orcis/trans isomers). According to the disclosure, the chemical structuresdepicted herein, and therefore the compounds of the disclosure,encompass all of the corresponding stereoisomers, that is, both thestereomerically pure form (e.g., geometrically pure, enantiomericallypure, or diastereomerically pure) and enantiomeric and stereoisomericmixtures, e.g., racemates. Enantiomeric and stereoisomeric mixtures ofcompounds of the disclosure can typically be resolved into theircomponent enantiomers or stereoisomers by well-known methods, such aschiral-phase gas chromatography, chiral-phase high performance liquidchromatography, crystallizing the compound as a chiral salt complex, orcrystallizing the compound in a chiral solvent. Enantiomers andstereoisomers can also be obtained from stereomerically orenantiomerically pure intermediates, reagents, and catalysts bywell-known asymmetric synthetic methods.

Linker: As used herein, a “linker” (including a membrane linker, asubunit linker, and a heterologous polypeptide linker as referred toherein) refers to a group of atoms, e.g., 10-1,000 atoms, and can becomprised of the atoms or groups such as, but not limited to, carbon,amino, alkylamino, oxygen, sulfur, sulfoxide, sulfonyl, carbonyl, andimine. The linker can be attached to a modified nucleoside or nucleotideon the nucleobase or sugar moiety at a first end, and to a payload,e.g., a detectable or therapeutic agent, at a second end. The linker canbe of sufficient length as to not interfere with incorporation into anucleic acid sequence. The linker can be used for any useful purpose,such as to form polynucleotide multimers (e.g., through linkage of twoor more chimeric polynucleotides molecules or IVT polynucleotides) orpolynucleotides conjugates, as well as to administer a payload, asdescribed herein. Examples of chemical groups that can be incorporatedinto the linker include, but are not limited to, alkyl, alkenyl,alkynyl, amido, amino, ether, thioether, ester, alkylene,heteroalkylene, aryl, or heterocyclyl, each of which can be optionallysubstituted, as described herein. Examples of linkers include, but arenot limited to, unsaturated alkanes, polyethylene glycols (e.g.,ethylene or propylene glycol monomeric units, e.g., diethylene glycol,dipropylene glycol, triethylene glycol, tripropylene glycol,tetraethylene glycol, or tetraethylene glycol), and dextran polymers andderivatives thereof. Other examples include, but are not limited to,cleavable moieties within the linker, such as, for example, a disulfidebond (—S—S—) or an azo bond (—N═N—), which can be cleaved using areducing agent or photolysis. Non-limiting examples of a selectivelycleavable bond include an amido bond can be cleaved for example by theuse of tris(2-carboxyethyl)phosphine (TCEP), or other reducing agents,and/or photolysis, as well as an ester bond can be cleaved for exampleby acidic or basic hydrolysis.

Methods of Administration: As used herein, “methods of administration”may include intravenous, intramuscular, intradermal, subcutaneous, orother methods of delivering a composition to a subject. A method ofadministration may be selected to target delivery (e.g., to specificallydeliver) to a specific region or system of a body.

Modified: As used herein “modified” refers to a changed state orstructure of a molecule of the disclosure. Molecules can be modified inmany ways including chemically, structurally, and functionally. In someembodiments, the mRNA molecules of the present disclosure are modifiedby the introduction of non-natural nucleosides and/or nucleotides, e.g.,as it relates to the natural ribonucleotides A, U, G, and C.Noncanonical nucleotides such as the cap structures are not considered“modified” although they differ from the chemical structure of the A, C,G, U ribonucleotides.

Mucus: As used herein, “mucus” refers to the natural substance that isviscous and comprises mucin glycoproteins.

Nanoparticle Composition: As used herein, a “nanoparticle composition”is a composition comprising one or more lipids. Nanoparticlecompositions are typically sized on the order of micrometers or smallerand may include a lipid bilayer. Nanoparticle compositions encompasslipid nanoparticles (LNPs), liposomes (e.g., lipid vesicles), andlipoplexes. For example, a nanoparticle composition may be a liposomehaving a lipid bilayer with a diameter of 500 nm or less.

Naturally occurring: As used herein, “naturally occurring” meansexisting in nature without artificial aid.

Non-human vertebrate: As used herein, a “non-human vertebrate” includesall vertebrates except Homo sapiens, including wild and domesticatedspecies. Examples of non-human vertebrates include, but are not limitedto, mammals, such as alpaca, banteng, bison, camel, cat, cattle, deer,dog, donkey, gayal, goat, guinea pig, horse, llama, mule, pig, rabbit,reindeer, sheep water buffalo, and yak.

Nucleic acid sequence: The terms “nucleic acid sequence,” “nucleotidesequence,” or “polynucleotide sequence” are used interchangeably andrefer to a contiguous nucleic acid sequence. The sequence can be eithersingle stranded or double stranded DNA or RNA, e.g., an mRNA.

The term “nucleic acid,” in its broadest sense, includes any compoundand/or substance that comprises a polymer of nucleotides. These polymersare often referred to as polynucleotides. Exemplary nucleic acids orpolynucleotides of the disclosure include, but are not limited to,ribonucleic acids (RNAs), deoxyribonucleic acids (DNAs), threose nucleicacids (TNAs), glycol nucleic acids (GNAs), peptide nucleic acids (PNAs),locked nucleic acids (LNAs, including LNA having a β-D-riboconfiguration, α-LNA having an α-L-ribo configuration (a diastereomer ofLNA), 2′-amino-LNA having a 2′-amino functionalization, and2′-amino-α-LNA having a 2′-amino functionalization), ethylene nucleicacids (ENA), cyclohexenyl nucleic acids (CeNA) or hybrids orcombinations thereof.

The phrase “nucleotide sequence encoding” refers to the nucleic acid(e.g., an mRNA or DNA molecule) coding sequence which encodes apolypeptide. The coding sequence can further include initiation andtermination signals operably linked to regulatory elements including apromoter and polyadenylation signal capable of directing expression inthe cells of an individual or mammal to which the nucleic acid isadministered. The coding sequence can further include sequences thatencode signal peptides.

Off-target: As used herein, “off target” refers to any unintended effecton any one or more target, gene, or cellular transcript.

Open reading frame: As used herein, “open reading frame” or “ORF” refersto a sequence which does not contain a stop codon in a given readingframe.

Operably linked: As used herein, the phrase “operably linked” refers toa functional connection between two or more molecules, constructs,transcripts, entities, moieties or the like.

Optionally substituted: Herein a phrase of the form “optionallysubstituted X” (e.g., optionally substituted alkyl) is intended to beequivalent to “X, wherein X is optionally substituted” (e.g., “alkyl,wherein said alkyl is optionally substituted”). It is not intended tomean that the feature “X” (e.g., alkyl) per se is optional.

Part: As used herein, a “part” or “region” of a polynucleotide isdefined as any portion of the polynucleotide that is less than theentire length of the polynucleotide.

Patient: As used herein, “patient” refers to a subject who may seek orbe in need of treatment, requires treatment, is receiving treatment,will receive treatment, or a subject who is under care by a trainedprofessional for a particular disease or condition.

Pharmaceutically acceptable: The phrase “pharmaceutically acceptable” isemployed herein to refer to those compounds, materials, compositions,and/or dosage forms that are, within the scope of sound medicaljudgment, suitable for use in contact with the tissues of human beingsand animals without excessive toxicity, irritation, allergic response,or other problem or complication, commensurate with a reasonablebenefit/risk ratio.

Pharmaceutically acceptable excipients: The phrase “pharmaceuticallyacceptable excipient,” as used herein, refers any ingredient other thanthe compounds described herein (for example, a vehicle capable ofsuspending or dissolving the active compound) and having the propertiesof being substantially nontoxic and non-inflammatory in a patient.Excipients can include, for example: antiadherents, antioxidants,binders, coatings, compression aids, disintegrants, dyes (colors),emollients, emulsifiers, fillers (diluents), film formers or coatings,flavors, fragrances, glidants (flow enhancers), lubricants,preservatives, printing inks, sorbents, suspensing or dispersing agents,sweeteners, and waters of hydration. Exemplary excipients include, butare not limited to: butylated hydroxytoluene (BHT), calcium carbonate,calcium phosphate (dibasic), calcium stearate, croscarmellose,crosslinked polyvinyl pyrrolidone, citric acid, crospovidone, cysteine,ethylcellulose, gelatin, hydroxypropyl cellulose, hydroxypropylmethylcellulose, lactose, magnesium stearate, maltitol, mannitol,methionine, methylcellulose, methyl paraben, microcrystalline cellulose,polyethylene glycol, polyvinyl pyrrolidone, povidone, pregelatinizedstarch, propyl paraben, retinyl palmitate, shellac, silicon dioxide,sodium carboxymethyl cellulose, sodium citrate, sodium starch glycolate,sorbitol, starch (corn), stearic acid, sucrose, talc, titanium dioxide,vitamin A, vitamin E, vitamin C, and xylitol.

Pharmaceutically acceptable salts: The present disclosure also includespharmaceutically acceptable salts of the compounds described herein. Asused herein, “pharmaceutically acceptable salts” refers to derivativesof the disclosed compounds wherein the parent compound is modified byconverting an existing acid or base moiety to its salt form (e.g., byreacting the free base group with a suitable organic acid). Examples ofpharmaceutically acceptable salts include, but are not limited to,mineral or organic acid salts of basic residues such as amines; alkalior organic salts of acidic residues such as carboxylic acids; and thelike. Representative acid addition salts include acetate, acetic acid,adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzenesulfonic acid, benzoate, bisulfate, borate, butyrate, camphorate,camphorsulfonate, citrate, cyclopentanepropionate, digluconate,dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate,glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide,hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate,lactate, laurate, lauryl sulfate, malate, maleate, malonate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate,oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate,phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate,tartrate, thiocyanate, toluenesulfonate, undecanoate, valerate salts,and the like. Representative alkali or alkaline earth metal saltsinclude sodium, lithium, potassium, calcium, magnesium, and the like, aswell as nontoxic ammonium, quaternary ammonium, and amine cations,including, but not limited to ammonium, tetramethylammonium,tetraethylammonium, methylamine, dimethylamine, trimethylamine,triethylamine, ethylamine, and the like. The pharmaceutically acceptablesalts of the present disclosure include the conventional non-toxic saltsof the parent compound formed, for example, from non-toxic inorganic ororganic acids. The pharmaceutically acceptable salts of the presentdisclosure can be synthesized from the parent compound that contains abasic or acidic moiety by conventional chemical methods. Generally, suchsalts can be prepared by reacting the free acid or base forms of thesecompounds with a stoichiometric amount of the appropriate base or acidin water or in an organic solvent, or in a mixture of the two;generally, nonaqueous media like ether, ethyl acetate, ethanol,isopropanol, or acetonitrile are used. Lists of suitable salts are foundin Remington's Pharmaceutical Sciences, 17^(th) ed., Mack PublishingCompany, Easton, Pa., 1985, p. 1418, Pharmaceutical Salts: Properties,Selection, and Use, P. H. Stahl and C. G. Wermuth (eds.), Wiley-VCH,2008, and Berge et al., Journal of Pharmaceutical Science, 66, 1-19(1977), each of which is incorporated herein by reference in itsentirety.

Pharmaceutically acceptable solvate: The term “pharmaceuticallyacceptable solvate,” as used herein, means a compound of the disclosurewherein molecules of a suitable solvent are incorporated in the crystallattice. A suitable solvent is physiologically tolerable at the dosageadministered. For example, solvates can be prepared by crystallization,recrystallization, or precipitation from a solution that includesorganic solvents, water, or a mixture thereof. Examples of suitablesolvents are ethanol, water (for example, mono-, di-, and tri-hydrates),N-methylpyrrolidinone (NMP), dimethyl sulfoxide (DMSO),N,N′-dimethylformamide (DMF), N,N′-dimethylacetamide (DMAC),1,3-dimethyl-2-imidazolidinone (DMEU),1,3-dimethyl-3,4,5,6-tetrahydro-2-(1H)-pyrimidinone (DMPU), acetonitrile(ACN), propylene glycol, ethyl acetate, benzyl alcohol, 2-pyrrolidone,benzyl benzoate, and the like. When water is the solvent, the solvate isreferred to as a “hydrate.”

Pharmacokinetic: As used herein, “pharmacokinetic” refers to any one ormore properties of a molecule or compound as it relates to thedetermination of the fate of substances administered to a livingorganism. Pharmacokinetics is divided into several areas including theextent and rate of absorption, distribution, metabolism and excretion.This is commonly referred to as ADME where: (A) Absorption is theprocess of a substance entering the blood circulation; (D) Distributionis the dispersion or dissemination of substances throughout the fluidsand tissues of the body; (M) Metabolism (or Biotransformation) is theirreversible transformation of parent compounds into daughtermetabolites; and (E) Excretion (or Elimination) refers to theelimination of the substances from the body. In rare cases, some drugsirreversibly accumulate in body tissue.

Physicochemical: As used herein, “physicochemical” means of or relatingto a physical and/or chemical property.

Polynucleotide: The term “polynucleotide” as used herein refers topolymers of nucleotides of any length, including ribonucleotides,deoxyribonucleotides, analogs thereof, or mixtures thereof. This termrefers to the primary structure of the molecule. Thus, the term includestriple-, double- and single-stranded deoxyribonucleic acid (“DNA”), aswell as triple-, double- and single-stranded ribonucleic acid (“RNA”).It also includes modified, for example by alkylation, and/or by capping,and unmodified forms of the polynucleotide. More particularly, the term“polynucleotide” includes polydeoxyribonucleotides (containing2-deoxy-D-ribose), polyribonucleotides (containing D-ribose), includingtRNA, rRNA, hRNA, siRNA and mRNA, whether spliced or unspliced, anyother type of polynucleotide which is an N- or C-glycoside of a purineor pyrimidine base, and other polymers containing normucleotidicbackbones, for example, polyamide (e.g., peptide nucleic acids “PNAs”)and polymorpholino polymers, and other synthetic sequence-specificnucleic acid polymers providing that the polymers contain nucleobases ina configuration which allows for base pairing and base stacking, such asis found in DNA and RNA. In particular aspects, the polynucleotidecomprises an mRNA. In other aspect, the mRNA is a synthetic mRNA. Insome aspects, the synthetic mRNA comprises at least one unnaturalnucleobase. In some aspects, all nucleobases of a certain class havebeen replaced with unnatural nucleobases (e.g., all uridines in apolynucleotide disclosed herein can be replaced with an unnaturalnucleobase, e.g., 5-methoxyuridine). In some aspects, the polynucleotide(e.g., a synthetic RNA or a synthetic DNA) comprises only naturalnucleobases, i.e., A (adenosine), G (guanosine), C (cytidine), and T(thymidine) in the case of a synthetic DNA, or A, C, G, and U (uridine)in the case of a synthetic RNA.

The skilled artisan will appreciate that the T bases in the codon mapsdisclosed herein are present in DNA, whereas the T bases would bereplaced by U bases in corresponding RNAs. For example, acodon-nucleotide sequence disclosed herein in DNA form, e.g., a vectoror an in-vitro translation (IVT) template, would have its T basestranscribed as U based in its corresponding transcribed mRNA. In thisrespect, both codon-optimized DNA sequences (comprising T) and theircorresponding mRNA sequences (comprising U) are consideredcodon-optimized nucleotide sequence of the present disclosure. A skilledartisan would also understand that equivalent codon-maps can begenerated by replaced one or more bases with non-natural bases. Thus,e.g., a TTC codon (DNA map) would correspond to a UUC codon (RNA map),which in turn would correspond to a ΨΨC codon (RNA map in which U hasbeen replaced with pseudouridine).

Standard A-T and G-C base pairs form under conditions which allow theformation of hydrogen bonds between the N3-H and C4-oxy of thymidine andthe N1 and C6-NH2, respectively, of adenosine and between the C2-oxy, N3and C4-NH2, of cytidine and the C2-NH2, N’—H and C6-oxy, respectively,of guanosine. Thus, for example, guanosine(2-amino-6-oxy-9-β-D-ribofuranosyl-purine) can be modified to formisoguanosine (2-oxy-6-amino-9-β-D-ribofuranosyl-purine). Suchmodification results in a nucleoside base which will no longereffectively form a standard base pair with cytosine. However,modification of cytosine (1-β-D-ribofuranosyl-2-oxy-4-amino-pyrimidine)to form isocytosine (113-D-ribofuranosyl-2-amino-4-oxy-pyrimidine-)results in a modified nucleotide which will not effectively base pairwith guanosine but will form a base pair with isoguanosine (U.S. Pat.No. 5,681,702 to Collins et al.). Isocytosine is available from SigmaChemical Co. (St. Louis, Mo.); isocytidine can be prepared by the methoddescribed by Switzer et al. (1993) Biochemistry 32:10489-10496 andreferences cited therein; 2′-deoxy-5-methyl-isocytidine can be preparedby the method of Tor et al., 1993, J. Am. Chem. Soc. 115:4461-4467 andreferences cited therein; and isoguanine nucleotides can be preparedusing the method described by Switzer et al., 1993, supra, and Mantschet al., 1993, Biochem. 14:5593-5601, or by the method described in U.S.Pat. No. 5,780,610 to Collins et al. Other nonnatural base pairs can besynthesized by the method described in Piccirilli et al., 1990, Nature343:33-37, for the synthesis of 2,6-diaminopyrimidine and its complement(1-methylpyrazolo-[4,3]pyrimidine-5,7-(4H,6H)-dione. Other such modifiednucleotide units which form unique base pairs are known, such as thosedescribed in Leach et al. (1992) J. Am. Chem. Soc. 114:3675-3683 andSwitzer et al., supra.

Polypeptide: The terms “polypeptide,” “peptide,” and “protein” are usedinterchangeably herein to refer to polymers of amino acids of anylength. The polymer can comprise modified amino acids. The terms alsoencompass an amino acid polymer that has been modified naturally or byintervention; for example, disulfide bond formation, glycosylation,lipidation, acetylation, phosphorylation, or any other manipulation ormodification, such as conjugation with a labeling component. Alsoincluded within the definition are, for example, polypeptides containingone or more analogs of an amino acid (including, for example, unnaturalamino acids such as homocysteine, ornithine, p-acetylphenylalanine,D-amino acids, and creatine), as well as other modifications known inthe art.

The term, as used herein, refers to proteins, polypeptides, and peptidesof any size, structure, or function. Polypeptides include encodedpolynucleotide products, naturally occurring polypeptides, syntheticpolypeptides, homologs, orthologs, paralogs, fragments and otherequivalents, variants, and analogs of the foregoing. A polypeptide canbe a monomer or can be a multi-molecular complex such as a dimer, trimeror tetramer. They can also comprise single chain or multichainpolypeptides. Most commonly disulfide linkages are found in multichainpolypeptides. The term polypeptide can also apply to amino acid polymersin which one or more amino acid residues are an artificial chemicalanalogue of a corresponding naturally occurring amino acid. In someembodiments, a “peptide” can be less than or equal to 50 amino acidslong, e.g., about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 amino acidslong.

Polypeptide variant: As used herein, the term “polypeptide variant”refers to molecules that differ in their amino acid sequence from anative or reference sequence. The amino acid sequence variants canpossess substitutions, deletions, and/or insertions at certain positionswithin the amino acid sequence, as compared to a native or referencesequence. Ordinarily, variants will possess at least about 50% identity,at least about 60% identity, at least about 70% identity, at least about80% identity, at least about 90% identity, at least about 95% identity,at least about 99% identity to a native or reference sequence. In someembodiments, they will be at least about 80%, or at least about 90%identical to a native or reference sequence.

Polypeptide per unit drug (PUD): As used herein, a PUD or product perunit drug, is defined as a subdivided portion of total daily dose,usually 1 mg, pg, kg, etc., of a product (such as a polypeptide) asmeasured in body fluid or tissue, usually defined in concentration suchas pmol/mL, mmol/mL, etc. divided by the measure in the body fluid.

Preventing: As used herein, the term “preventing” refers to partially orcompletely delaying onset of an infection, disease, disorder and/orcondition; partially or completely delaying onset of one or moresymptoms, features, or clinical manifestations of a particularinfection, disease, disorder, and/or condition; partially or completelydelaying onset of one or more symptoms, features, or manifestations of aparticular infection, disease, disorder, and/or condition; partially orcompletely delaying progression from an infection, a particular disease,disorder and/or condition; and/or decreasing the risk of developingpathology associated with the infection, the disease, disorder, and/orcondition.

Proliferate: As used herein, the term “proliferate” means to grow,expand or increase or cause to grow, expand or increase rapidly.“Proliferative” means having the ability to proliferate.“Anti-proliferative” means having properties counter to or inapposite toproliferative properties. Prophylactic: As used herein, “prophylactic”refers to a therapeutic or course of action used to prevent the spreadof disease.

Prophylaxis: As used herein, a “prophylaxis” refers to a measure takento maintain health and prevent the spread of disease. An “immuneprophylaxis” refers to a measure to produce active or passive immunityto prevent the spread of disease.

Protein cleavage site: As used herein, “protein cleavage site” refers toa site where controlled cleavage of the amino acid chain can beaccomplished by chemical, enzymatic or photochemical means.

Protein cleavage signal: As used herein “protein cleavage signal” refersto at least one amino acid that flags or marks a polypeptide forcleavage.

Protein of interest: As used herein, the terms “proteins of interest” or“desired proteins” include those provided herein and fragments, mutants,variants, and alterations thereof.

Proximal: As used herein, the term “proximal” means situated nearer tothe center or to a point or region of interest.

Pseudouridine: As used herein, pseudouridine (w) refers to theC-glycoside isomer of the nucleoside uridine. A “pseudouridine analog”is any modification, variant, isoform or derivative of pseudouridine.For example, pseudouridine analogs include but are not limited to1-carboxymethyl-pseudouridine, 1-propynyl-pseudouridine,1-taurinomethyl-pseudouridine, 1-taurinomethyl-4-thio-pseudouridine,1-methylpseudouridine (m′w), 1-methyl-4-thio-pseudouridine (m's⁴ψ),4-thio-1-methyl-pseudouridine, 3-methyl-pseudouridine (m³ψ),2-thio-1-methyl-pseudouridine, 1-methyl-1-deaza-pseudouridine,2-thio-1-methyl-1-deaza-pseudouridine, dihydropseudouridine,2-thio-dihydropseudouridine, 2-methoxyuridine, 2-methoxy-4-thio-uridine,4-methoxy-pseudouridine, 4-methoxy-2-thio-pseudouridine,N1-methyl-pseudouridine,1-methyl-3-(3-amino-3-carboxypropyl)pseudouridine (acp³ ψ), and2′-O-methyl-pseudouridine (ψm).

Purified: As used herein, “purify,” “purified,” “purification” means tomake substantially pure or clear from unwanted components, materialdefilement, admixture or imperfection.

Reference Nucleic Acid Sequence: The term “reference nucleic acidsequence” or “reference nucleic acid” or “reference nucleotide sequence”or “reference sequence” refers to a starting nucleic acid sequence(e.g., a RNA, e.g., an mRNA sequence) that can be sequence optimized. Insome embodiments, the reference nucleic acid sequence is a wild typenucleic acid sequence, a fragment or a variant thereof. In someembodiments, the reference nucleic acid sequence is a previouslysequence optimized nucleic acid sequence.

Salts: In some aspects, the pharmaceutical composition for intratumoraldelivery disclosed herein and comprises salts of some of their lipidconstituents. The term “salt” includes any anionic and cationic complex.Non-limiting examples of anions include inorganic and organic anions,e.g., fluoride, chloride, bromide, iodide, oxalate (e.g., hemioxalate),phosphate, phosphonate, hydrogen phosphate, dihydrogen phosphate, oxide,carbonate, bicarbonate, nitrate, nitrite, nitride, bisulfite, sulfide,sulfite, bisulfate, sulfate, thiosulfate, hydrogen sulfate, borate,formate, acetate, benzoate, citrate, tartrate, lactate, acrylate,polyacrylate, fumarate, maleate, itaconate, glycolate, gluconate,malate, mandelate, tiglate, ascorbate, salicylate, polymethacrylate,perchlorate, chlorate, chlorite, hypochlorite, bromate, hypobromite,iodate, an alkylsulfonate, an arylsulfonate, arsenate, arsenite,chromate, dichromate, cyanide, cyanate, thiocyanate, hydroxide,peroxide, permanganate, and mixtures thereof.

Sample: As used herein, the term “sample” or “biological sample” refersto a subset of its tissues, cells or component parts (e.g., body fluids,including but not limited to blood, mucus, lymphatic fluid, synovialfluid, cerebrospinal fluid, saliva, amniotic fluid, amniotic cord blood,urine, vaginal fluid and semen). A sample further can include ahomogenate, lysate or extract prepared from a whole organism or a subsetof its tissues, cells or component parts, or a fraction or portionthereof, including but not limited to, for example, plasma, serum,spinal fluid, lymph fluid, the external sections of the skin,respiratory, intestinal, and genitourinary tracts, tears, saliva, milk,blood cells, tumors, organs. A sample further refers to a medium, suchas a nutrient broth or gel, which may contain cellular components, suchas proteins or nucleic acid molecule.

Signal Sequence: As used herein, the phrases “signal sequence,” “signalpeptide,” and “transit peptide” are used interchangeably and refer to asequence that can direct the transport or localization of a protein to acertain organelle, cell compartment, or extracellular export. The termencompasses both the signal sequence polypeptide and the nucleic acidsequence encoding the signal sequence. Thus, references to a signalsequence in the context of a nucleic acid refer in fact to the nucleicacid sequence encoding the signal sequence polypeptide.

Signal transduction pathway: A “signal transduction pathway” refers tothe biochemical relationship between a variety of signal transductionmolecules that play a role in the transmission of a signal from oneportion of a cell to another portion of a cell. As used herein, thephrase “cell surface receptor” includes, for example, molecules andcomplexes of molecules capable of receiving a signal and thetransmission of such a signal across the plasma membrane of a cell.

Similarity: As used herein, the term “similarity” refers to the overallrelatedness between polymeric molecules, e.g. between polynucleotidemolecules (e.g. DNA molecules and/or RNA molecules) and/or betweenpolypeptide molecules. Calculation of percent similarity of polymericmolecules to one another can be performed in the same manner as acalculation of percent identity, except that calculation of percentsimilarity takes into account conservative substitutions as isunderstood in the art.

Single unit dose: As used herein, a “single unit dose” is a dose of anytherapeutic administered in one dose/at one time/single route/singlepoint of contact, i.e., single administration event.

Split dose: As used herein, a “split dose” is the division of singleunit dose or total daily dose into two or more doses.

Specific delivery: As used herein, the term “specific delivery,”“specifically deliver,” or “specifically delivering” means delivery ofmore (e.g., at least 1.5 fold more, at least 2-fold more, at least3-fold more, at least 4-fold more, at least 5-fold more, at least 6-foldmore, at least 7-fold more, at least 8-fold more, at least 9-fold more,at least 10-fold more) of a polynucleotide by a nanoparticle to a targettissue of interest (e.g., mammalian liver) compared to an off-targettissue (e.g., mammalian spleen). The level of delivery of a nanoparticleto a particular tissue may be measured by comparing the amount ofprotein produced in a tissue to the weight of said tissue, comparing theamount of polynucleotide in a tissue to the weight of said tissue,comparing the amount of protein produced in a tissue to the amount oftotal protein in said tissue, or comparing the amount of polynucleotidein a tissue to the amount of total polynucleotide in said tissue. Forexample, for renovascular targeting, a polynucleotide is specificallyprovided to a mammalian kidney as compared to the liver and spleen if1.5, 2-fold, 3-fold, 5-fold, 10-fold, 15 fold, or 20 fold morepolynucleotide per 1 g of tissue is delivered to a kidney compared tothat delivered to the liver or spleen following systemic administrationof the polynucleotide. It will be understood that the ability of ananoparticle to specifically deliver to a target tissue need not bedetermined in a subject being treated, it may be determined in asurrogate such as an animal model (e.g., a rat model).

Stable: As used herein “stable” refers to a compound that issufficiently robust to survive isolation to a useful degree of purityfrom a reaction mixture, and in some cases capable of formulation intoan efficacious therapeutic agent.

Stabilized: As used herein, the term “stabilize,” “stabilized,”“stabilized region” means to make or become stable.

Stereoisomer: As used herein, the term “stereoisomer” refers to allpossible different isomeric as well as conformational forms that acompound may possess (e.g., a compound of any formula described herein),in particular all possible stereochemically and conformationallyisomeric forms, all diastereomers, enantiomers and/or conformers of thebasic molecular structure. Some compounds of the present disclosure mayexist in different tautomeric forms, all of the latter being includedwithin the scope of the present disclosure.

Subject: By “subject” or “individual” or “animal” or “patient” or“mammal,” is meant any subject, particularly a mammalian subject, forwhom diagnosis, prognosis, or therapy is desired. Mammalian subjectsinclude, but are not limited to, humans, domestic animals, farm animals,zoo animals, sport animals, pet animals such as dogs, cats, guinea pigs,rabbits, rats, mice, horses, cattle, cows; primates such as apes,monkeys, orangutans, and chimpanzees; canids such as dogs and wolves;felids such as cats, lions, and tigers; equids such as horses, donkeys,and zebras; bears, food animals such as cows, pigs, and sheep; ungulatessuch as deer and giraffes; rodents such as mice, rats, hamsters andguinea pigs; and so on. In certain embodiments, the mammal is a humansubject. In other embodiments, a subject is a human patient. In aparticular embodiment, a subject is a human patient in need oftreatment.

Substantially: As used herein, the term “substantially” refers to thequalitative condition of exhibiting total or near-total extent or degreeof a characteristic or property of interest. One of ordinary skill inthe biological arts will understand that biological and chemicalcharacteristics rarely, if ever, go to completion and/or proceed tocompleteness or achieve or avoid an absolute result. The term“substantially” is therefore used herein to capture the potential lackof completeness inherent in many biological and chemicalcharacteristics.

Substantially equal: As used herein as it relates to time differencesbetween doses, the term means plus/minus 2%.

Substantially simultaneous: As used herein and as it relates toplurality of doses, the term means within 2 seconds.

Suffering from: An individual who is “suffering from” a disease,disorder, and/or condition has been diagnosed with or displays one ormore symptoms of the disease, disorder, and/or condition.

Susceptible to: An individual who is “susceptible to” a disease,disorder, and/or condition has not been diagnosed with and/or may notexhibit symptoms of the disease, disorder, and/or condition but harborsa propensity to develop a disease or its symptoms. In some embodiments,an individual who is susceptible to a disease, disorder, and/orcondition (for example, cancer) can be characterized by one or more ofthe following: (1) a genetic mutation associated with development of thedisease, disorder, and/or condition; (2) a genetic polymorphismassociated with development of the disease, disorder, and/or condition;(3) increased and/or decreased expression and/or activity of a proteinand/or nucleic acid associated with the disease, disorder, and/orcondition; (4) habits and/or lifestyles associated with development ofthe disease, disorder, and/or condition; (5) a family history of thedisease, disorder, and/or condition; and (6) exposure to and/orinfection with a microbe associated with development of the disease,disorder, and/or condition. In some embodiments, an individual who issusceptible to a disease, disorder, and/or condition will develop thedisease, disorder, and/or condition. In some embodiments, an individualwho is susceptible to a disease, disorder, and/or condition will notdevelop the disease, disorder, and/or condition.

Sustained release: As used herein, the term “sustained release” refersto a pharmaceutical composition or compound release profile thatconforms to a release rate over a specific period of time.

Synthetic: The term “synthetic” means produced, prepared, and/ormanufactured by the hand of man. Synthesis of polynucleotides or othermolecules of the present disclosure can be chemical or enzymatic.

Targeted Cells: As used herein, “targeted cells” refers to any one ormore cells of interest. The cells can be found in vitro, in vivo, insitu or in the tissue or organ of an organism. The organism can be ananimal, for example a mammal, a human, a subject or a patient.

Target tissue: As used herein “target tissue” refers to any one or moretissue types of interest in which the delivery of a polynucleotide wouldresult in a desired biological and/or pharmacological effect. Examplesof target tissues of interest include specific tissues, organs, andsystems or groups thereof. In particular applications, a target tissuemay be a kidney, a lung, a spleen, vascular endothelium in vessels(e.g., intra-coronary or intra-femoral), or tumor tissue (e.g., viaintratumoral injection). An “off-target tissue” refers to any one ormore tissue types in which the expression of the encoded protein doesnot result in a desired biological and/or pharmacological effect. Inparticular applications, off-target tissues may include the liver andthe spleen.

The presence of a therapeutic agent in an off-target issue can be theresult of: (i) leakage of a polynucleotide from the administration siteto peripheral tissue or distant off-target tissue (e.g., liver) viadiffusion or through the bloodstream (e.g., a polynucleotide intended toexpress a polypeptide in a certain tissue would reach the liver and thepolypeptide would be expressed in the liver); or (ii) leakage of anpolypeptide after administration of a polynucleotide encoding suchpolypeptide to peripheral tissue or distant off-target tissue (e.g.,liver) via diffusion or through the bloodstream (e.g., a polynucleotidewould expressed a polypeptide in the target tissue, and the polypeptidewould diffuse to peripheral tissue).

Targeting sequence: As used herein, the phrase “targeting sequence”refers to a sequence that can direct the transport or localization of aprotein or polypeptide.

Terminus: As used herein the terms “termini” or “terminus,” whenreferring to polypeptides, refers to an extremity of a peptide orpolypeptide. Such extremity is not limited only to the first or finalsite of the peptide or polypeptide but can include additional aminoacids in the terminal regions. The polypeptide based molecules of thedisclosure can be characterized as having both an N-terminus (terminatedby an amino acid with a free amino group (NH2)) and a C-terminus(terminated by an amino acid with a free carboxyl group (COOH)).Proteins of the disclosure are in some cases made up of multiplepolypeptide chains brought together by disulfide bonds or bynon-covalent forces (multimers, oligomers). These sorts of proteins willhave multiple N- and C-termini. Alternatively, the termini of thepolypeptides can be modified such that they begin or end, as the casecan be, with a non-polypeptide based moiety such as an organicconjugate.

Therapeutic Agent: The term “therapeutic agent” refers to an agent that,when administered to a subject, has a therapeutic, diagnostic, and/orprophylactic effect and/or elicits a desired biological and/orpharmacological effect. For example, in some embodiments, an mRNAencoding a tethered IL-12 polypeptide as disclosed herein can be atherapeutic agent.

Therapeutically effective amount: As used herein, the term“therapeutically effective amount” means an amount of an agent to bedelivered (e.g., nucleic acid, drug, therapeutic agent, diagnosticagent, prophylactic agent, etc.) that is sufficient, when administeredto a subject suffering from or susceptible to an infection, disease,disorder, and/or condition, to treat, improve symptoms of, diagnose,prevent, and/or delay the onset of the infection, disease, disorder,and/or condition.

Therapeutically effective outcome: As used herein, the term“therapeutically effective outcome” means an outcome that is sufficientin a subject suffering from or susceptible to an infection, disease,disorder, and/or condition, to treat, improve symptoms of, diagnose,prevent, and/or delay the onset of the infection, disease, disorder,and/or condition.

Total daily dose: As used herein, a “total daily dose” is an amountgiven or prescribed in 24 hour period. The total daily dose can beadministered as a single unit dose or a split dose.

Transmembrane domain: As used herein, the terms “transmembrane domain”,“TM” and “TMD” refer to the region of a polypeptide which crosses theplasma membrane of a cell.

Transcription factor: As used herein, the term “transcription factor”refers to a DNA-binding protein that regulates transcription of DNA intoRNA, for example, by activation or repression of transcription. Sometranscription factors effect regulation of transcription alone, whileothers act in concert with other proteins. Some transcription factor canboth activate and repress transcription under certain conditions. Ingeneral, transcription factors bind a specific target sequence orsequences highly similar to a specific consensus sequence in aregulatory region of a target gene. Transcription factors may regulatetranscription of a target gene alone or in a complex with othermolecules.

Transcription: As used herein, the term “transcription” refers tomethods to introduce exogenous nucleic acids into a cell. Methods oftransfection include, but are not limited to, chemical methods, physicaltreatments and cationic lipids or mixtures.

Transcription start site: As used herein, the term “transcription startsite” refers to a specific nucleotide in the sense strand of a DNAmolecule where transcription by an RNA polymerase initiates and thatcorresponds to the first nucleotide in the transcript. The transcriptionstart site is typically located downstream of a promoter, which is aregion of DNA that initiations transcription. For example, the T7 RNApolymerase initiates transcription at the underlined G in the promotersequence 5′ TAATACGACTCACTATAG 3′. The polymerase then transcribes usingthe opposite DNA strand as a template. In some embodiments, thetranscription start site for a T7 RNA polymerase is referred to as a “T7start site”. The first base in the transcript will be a G. The DNAcontacts made by T7 RNA polymerase have been mapped during binding andduring the subsequent initiation of transcription. The RNA polymerasealone protects 19 bases in a region from −21 to −3. Synthesis of thetrinucleotide r(GGG) expands the length of the sequence protected by theRNA polymerase and stabilizes the complex. The formation of ahexanucleotide mRNA, r(GGGAGA) further extends the protected region,stabilizes the complex, and results in increased transcriptionalefficiency (Ikeda and Richardson (1986) Proc Natl Acad Sci83:3614-3618). The sequence GGGAGA is referred to as a “T7 leadersequence”. Accordingly, in some embodiments, the mRNAs provided by thedisclosure comprise a 5′ UTR comprising a T7 leader sequence at the 5′end of the 5′ UTR. In some embodiments, the mRNA of the disclosurecomprises a 5′ UTR comprising a T7 leader sequence comprising thesequence GGGAGA at the 5′ end of the 5′ UTR. In some embodiments, themRNA of the disclosure comprises a 5′ UTR comprising a T7 leadersequence comprising the sequence GGGAAA at the 5′ end of the 5′ UTR. Insome embodiments, the mRNA comprises a 5′ UTR which does not comprise aT7 leader sequence.

Transfection: As used herein, “transfection” refers to the introductionof a polynucleotide into a cell wherein a polypeptide encoded by thepolynucleotide is expressed (e.g., mRNA) or the polypeptide modulates acellular function (e.g., siRNA, miRNA). As used herein, “expression” ofa nucleic acid sequence refers to translation of a polynucleotide (e.g.,an mRNA) into a polypeptide or protein and/or post-translationalmodification of a polypeptide or protein.

Treating, treatment, therapy: As used herein, the term “treating” or“treatment” or “therapy” refers to partially or completely alleviating,ameliorating, improving, relieving, delaying onset of, inhibitingprogression of, reducing severity of, and/or reducing incidence of oneor more symptoms or features of a disease, e.g., acute intermittentporphyria. For example, “treating” acute intermittent porphyria canrefer to diminishing symptoms associate with the disease, prolong thelifespan (increase the survival rate) of patients, reducing the severityof the disease, preventing or delaying the onset of the disease, etc.Treatment can be administered to a subject who does not exhibit signs ofa disease, disorder, and/or condition and/or to a subject who exhibitsonly early signs of a disease, disorder, and/or condition for thepurpose of decreasing the risk of developing pathology associated withthe disease, disorder, and/or condition.

Type I integral membrane protein: As used herein, the term “type Iintegral membrane protein” refers to an integral membrane protein (i.e.,proteins having at least one transmembrane domain that crosses the lipidbilayer) with its amino-terminus in the extracellular space andcomprising one alpha-helical transmembrane domain.

Unmodified: As used herein, “unmodified” refers to any substance,compound or molecule prior to being changed in some way. Unmodified can,but does not always, refer to the wild type or native form of abiomolecule. Molecules can undergo a series of modifications wherebyeach modified molecule can serve as the “unmodified” starting moleculefor a subsequent modification.

Uracil: Uracil is one of the four nucleobases in the nucleic acid ofRNA, and it is represented by the letter U. Uracil can be attached to aribose ring, or more specifically, a ribofuranose via a 13-N1-glycosidicbond to yield the nucleoside uridine. The nucleoside uridine is alsocommonly abbreviated according to the one letter code of its nucleobase,i.e., U. Thus, in the context of the present disclosure, when a monomerin a polynucleotide sequence is U, such U is designated interchangeablyas a “uracil” or a “uridine.”

Uridine Content: The terms “uridine content” or “uracil content” areinterchangeable and refer to the amount of uracil or uridine present ina certain nucleic acid sequence. Uridine content or uracil content canbe expressed as an absolute value (total number of uridine or uracil inthe sequence) or relative (uridine or uracil percentage respect to thetotal number of nucleobases in the nucleic acid sequence).

Uridine Modified Sequence: The terms “uridine-modified sequence” refersto a sequence optimized nucleic acid (e.g., a synthetic mRNA sequence)with a different overall or local uridine content (higher or loweruridine content) or with different uridine patterns (e.g., gradientdistribution or clustering) with respect to the uridine content and/oruridine patterns of a candidate nucleic acid sequence. In the content ofthe present disclosure, the terms “uridine-modified sequence” and“uracil-modified sequence” are considered equivalent andinterchangeable.

A “high uridine codon” is defined as a codon comprising two or threeuridines, a “low uridine codon” is defined as a codon comprising oneuridine, and a “no uridine codon” is a codon without any uridines. Insome embodiments, a uridine-modified sequence comprises substitutions ofhigh uridine codons with low uridine codons, substitutions of highuridine codons with no uridine codons, substitutions of low uridinecodons with high uridine codons, substitutions of low uridine codonswith no uridine codons, substitution of no uridine codons with lowuridine codons, substitutions of no uridine codons with high uridinecodons, and combinations thereof. In some embodiments, a high uridinecodon can be replaced with another high uridine codon. In someembodiments, a low uridine codon can be replaced with another lowuridine codon. In some embodiments, a no uridine codon can be replacedwith another no uridine codon. A uridine-modified sequence can beuridine enriched or uridine rarefied.

Uridine Enriched: As used herein, the terms “uridine enriched” andgrammatical variants refer to the increase in uridine content (expressedin absolute value or as a percentage value) in a sequence optimizednucleic acid (e.g., a synthetic mRNA sequence) with respect to theuridine content of the corresponding candidate nucleic acid sequence.Uridine enrichment can be implemented by substituting codons in thecandidate nucleic acid sequence with synonymous codons containing lessuridine nucleobases. Uridine enrichment can be global (i.e., relative tothe entire length of a candidate nucleic acid sequence) or local (i.e.,relative to a subsequence or region of a candidate nucleic acidsequence).

Uridine Rarefied: As used herein, the terms “uridine rarefied” andgrammatical variants refer to a decrease in uridine content (expressedin absolute value or as a percentage value) in a sequence optimizednucleic acid (e.g., a synthetic mRNA sequence) with respect to theuridine content of the corresponding candidate nucleic acid sequence.Uridine rarefication can be implemented by substituting codons in thecandidate nucleic acid sequence with synonymous codons containing lessuridine nucleobases. Uridine rarefication can be global (i.e., relativeto the entire length of a candidate nucleic acid sequence) or local(i.e., relative to a subsequence or region of a candidate nucleic acidsequence).

Variant: The term variant as used in present disclosure refers to bothnatural variants (e.g., polymorphisms, isoforms, etc) and artificialvariants in which at least one amino acid residue in a native orstarting sequence (e.g., a wild type sequence) has been removed and adifferent amino acid inserted in its place at the same position. Thesevariants can be described as “substitutional variants.” Thesubstitutions can be single, where only one amino acid in the moleculehas been substituted, or they can be multiple, where two or more aminoacids have been substituted in the same molecule. If amino acids areinserted or deleted, the resulting variant would be an “insertionalvariant” or a “deletional variant” respectively.

The terms “invention” and “disclosure” can be used interchangeably whendescribing or used, for example, in the phrases “the present invention”or “the present disclosure.”

31. Other Embodiments

E1. A polynucleotide comprising an open reading frame (ORF) comprising:(a) a first nucleic acid sequence encoding an Interleukin-12 p40 subunit(IL-12B), (b) a second nucleic acid sequence encoding an Interleukin-12p35 subunit (IL-12A), and (c) a nucleic acid sequence encoding atransmembrane domain,

-   -   wherein the first nucleic acid sequence and the second nucleic        acid sequence are linked by a nucleic acid sequence encoding a        linker (“subunit linker”), and    -   wherein the nucleic acid sequence encoding the transmembrane        domain is linked to the first or second nucleic acid sequence by        a nucleic acid sequence encoding a linker (“transmembrane domain        linker”).

E2. The polynucleotide of embodiment 1, wherein the first nucleic acidsequence is located at the 5′ end of the subunit linker.

E3. The polynucleotide of embodiment 2, wherein the nucleic acidsequence encoding the transmembrane domain is located at the 3′ end ofthe transmembrane domain linker.

E4. The polynucleotide of any one of embodiments 1 to 3, wherein thepolynucleotide further comprises a nucleic acid sequence encoding asignal peptide.

E5. The polynucleotide of embodiment 4, wherein the nucleic acidsequence encoding the signal peptide is located at the 5′ end of thefirst nucleic acid sequence.

E6. The polynucleotide of any one of embodiments 1 to 5, wherein theIL12B has an amino acid sequence at least about 80%, at least about 90%,at least about 95%, at least 96%, at least 97%, at least 98%, at least99%, or 100% identical to amino acids 23 to 328 of SEQ ID NO: 48, andwherein the amino acid sequence has IL12B activity.

E7. The polynucleotide of any one of embodiments 1 to 6, wherein theIL12A has an amino acid sequence at least about 80%, at least about 90%,at least about 95%, at least 96%, at least 97%, at least 98%, at least99%, or 100% identical to amino acids 336 to 532 of SEQ ID NO: 48, andwherein the amino acid sequence has IL12A activity.

E8. The polynucleotide of any one of embodiments 4 to 7, wherein thesignal peptide comprises a sequence at least about 80%, at least about90%, at least about 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identical to amino acids 1 to 22 of SEQ ID NO: 48.

E9. The polynucleotide of any one of embodiments 1 to 8, wherein thesubunit linker is a Gly/Ser linker.

E10. The polynucleotide of any one of embodiments 1 to 9, wherein thetransmembrane domain linker is a Gly/Ser linker.

E11. The polynucleotide of embodiment 9 or embodiment 10, wherein theGly/Ser linker comprises (G_(n)S)_(m), wherein n is 1, 2 3, 4, 5, 6, 7,8, 9, 10, 15, or 20 and m is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, or 20.

E12. The polynucleotide of any one of embodiments 1-11, wherein thetransmembrane domain is a type I transmembrane domain.

E13. The polynucleotide of any one of embodiments 1-12, wherein thetransmembrane domain is a Cluster of Differentiation 8 (CD8)transmembrane domain or a Platelet-Derived Growth Factor Receptor(PDGF-R) transmembrane domain.

E14. The polynucleotide of any one of embodiments 1 to 13, wherein thepolynucleotide is DNA.

E15. The polynucleotide of any one of embodiments 1 to 13, wherein thepolynucleotide is RNA.

E16. The polynucleotide of embodiment 15, wherein the polynucleotide ismRNA.

E17. The polynucleotide of any one of embodiments 1 to 16, wherein thepolynucleotide comprises at least one chemically modified nucleobase.

E18. The polynucleotide of embodiment 17, wherein the at least onechemically modified nucleobase is selected from the group consisting ofpseudouracil (ψ), N1-methylpseudouracil (m1ψ), 2-thiouracil (s2U),4′-thiouracil, 5-methylcytosine, 2-thio-1-methyl-1-deaza-pseudouracil,2-thio-1-methyl-pseudouracil, 2-thio-5-aza-uracil,2-thio-dihydropseudouracil, 2-thio-dihydrouracil, 2-thio-pseudouracil,4-methoxy-2-thio-pseudouracil, 4-methoxy-pseudouracil,4-thio-1-methyl-pseudouracil, 4-thio-pseudouracil, 5-aza-uracil,dihydropseudouracil, 5-methyluracil, 5-methoxyuracil, 2′-O-methyluracil, 1-methyl-pseudouracil (m1ψ), 5-methoxy-uracil (mo5U),5-methyl-cytosine (m5C), α-thio-guanine, α-thio-adenine, 5-cyano uracil,4′-thio uracil, 7-deaza-adenine, 1-methyl-adenine (m1A),2-methyl-adenine (m2A), N6-methyl-adenine (m6A), and 2,6-Diaminopurine,(I), 1-methyl-inosine (mll), wyosine (imG), methylwyosine (mimG),7-deaza-guanine, 7-cyano-7-deaza-guanine (preQ0),7-aminomethyl-7-deaza-guanine (preQ1), 7-methyl-guanine (m7G),1-methyl-guanine (m1G), 8-oxo-guanine, 7-methyl-8-oxo-guanine, and twoor more combinations thereof.

E19. The polynucleotide of embodiment 17 or 18, wherein the nucleobasesin the polynucleotide are chemically modified by at least about 10%, atleast about 20%, at least about 30%, at least about 40%, at least about50%, at least about 60%, at least about 70%, at least about 80%, atleast about 90%, at least 95%, at least 99%, or 100%.

E20. The polynucleotide of any one of embodiments 17 to 19, wherein thechemically modified nucleobases are selected from the group consistingof uracil, adenine, cytosine, guanine, and any combination thereof.

E21. The polynucleotide of any one of embodiments 17 to 20, wherein theuracils, adenines, cytosines or guanines are chemically modified by atleast about 10%, at least about 20%, at least about 30%, at least about40%, at least about 50%, at least about 60%, at least about 70%, atleast about 80%, at least about 90%, at least 95%, at least 99%, or100%.

E22. The polynucleotide of any one of embodiments 1 to 21, wherein thepolynucleotide further comprises a nucleic acid sequence comprising amiRNA binding site.

E23. The polynucleotide of embodiment 22, wherein the miRNA binding sitebinds to miR-122.

E24. The polynucleotide of embodiment 22 or 23, wherein the miRNAbinding site binds to miR-122-3p or miR-122-5p.

E25. The polynucleotide of any one of embodiments 1 to 24, wherein thepolynucleotide further comprises a 5′ UTR.

E26. The polynucleotide of embodiment 25, wherein the 5′ UTR comprises anucleic acid sequence at least 90%, 95%, 96%, 97%, 98%, 99%, or 100%identical to any one of sequences disclosed herein.

E27. The polynucleotide of any one of embodiments 1 to 26, which furthercomprises a 3′ UTR. E28. The polynucleotide of embodiment 27, whereinthe 3′ UTR comprises a nucleic acid sequence at least about 90%, atleast about 95%, at least 96%, at least 97%, at least 98%, at least 99%,or 100% identical to any one of sequences disclosed herein.

E29. The polynucleotide of embodiment 27 or 28 wherein the miRNA bindingsite is located within the 3′ UTR.

E30. The polynucleotide of any one of embodiments 25 to 29, wherein the5′ UTR comprises a 5′ terminal cap.

E31. The polynucleotide of embodiment 30, wherein the 5′ terminal cap isa Cap0, Cap1, ARCA, inosine, N1-methyl-guanosine, 2′fluoro-guanosine,7-deaza-guanosine, 8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine,2-azidoguanosine, Cap2, Cap4, 5′ methylG cap, or an analog thereof. E32.The polynucleotide of any one of embodiments 1 to 31, wherein thepolynucleotide further comprises a poly-A region.

E33. The polynucleotide of embodiment 32, wherein the poly-A region isat least about 10, at least about 20, at least about 30, at least about40, at least about 50, at least about 60, at least about 70, at leastabout 80, or at least about 90 nucleotides in length.

E34. The polynucleotide of embodiment 32, wherein the poly-A region hasabout 10 to about 200 nucleotides in length, about 20 to about 180nucleotides in length, about 30 to about 160 nucleotides in length,about 40 to about 140 nucleotides in length, about 50 to about 120nucleotides in length, about 60 to about 100 nucleotides in length, orabout 80 to about 90 nucleotides in length.

E35. The polynucleotide of any one of embodiments 1 to 34, wherein thepolynucleotide has been transcribed in vitro (IVT).

E36. The polynucleotide of any one of embodiments 1 to 34, wherein thepolynucleotide is chimeric.

E37. The polynucleotide of any one of embodiments 1 to 34, wherein thepolynucleotide is circular.

E38. The polynucleotide of any one of embodiments 1 to 37, wherein theORF further comprises one or more nucleic acid sequences encoding one ormore heterologous polypeptides fused to the nucleic acid sequenceencoding the IL12B, the IL12A, or both.

E39. The polynucleotide of embodiment 38, wherein the one or moreheterologous polypeptides increase a pharmacokinetic property of theIL12A, the IL12B, or both.

E40. The polynucleotide of any one of embodiments 1 to 39, which issingle stranded.

E41. The polynucleotide of any one of embodiments 1 to 39, which isdouble stranded.

E42. The polynucleotide of any one of embodiments 1 to 41, wherein theIL12B is a variant, derivative, or a mutant having an IL12B activity.

E43. The polynucleotide of any one of embodiments 1 to 41, wherein theIL12A is a variant, derivative, or a mutant having an IL12A activity.

E44. A vector comprising the polynucleotide of any one of embodiments 1to 43.

E45. A composition comprising (i) the polynucleotide of any one ofembodiments 1 to 44 or the vector of embodiment 44, and (ii) a deliveryagent.

E46. The composition of embodiment 45, wherein the delivery agentcomprises a lipid nanoparticle.

E47. The composition of embodiment 46, wherein the lipid nanoparticlecomprises the compound of formula (I).

E48. The composition of any one of embodiments 45 to 47, wherein thedelivery agent further comprises a phospholipid.

E49. The composition of any one of embodiments 45 to 48, wherein thedelivery agent further comprises a structural lipid.

E50. The composition of embodiment 49, wherein the structural lipid ischolesterol.

E51. The composition of any one of embodiments 45 to 50, wherein thedelivery agent further comprises a PEG lipid.

E52. The composition of any one of embodiments 45 to 51, wherein thedelivery agent further comprises a quaternary amine compound.

E53. A method of reducing the size of a tumor or inhibiting growth of atumor in a subject in need thereof comprising administering thepolynucleotide of any one of embodiments 1 to 43, the vector ofembodiment 44, or the composition of any one of embodiments 45 to 52 inthe subject.

E54. The method of embodiment 53, wherein the polynucleotide, vector, orcomposition is administered subcutaneously, intravenously,intraperitoneally, or intratumorally.

E55. The method of embodiment 53 or 54, wherein the administrationtreats a cancer.

E56. The method of any one of embodiments 53 to 55, wherein thepolynucleotide is administered intratumorally to the subject.

E57. The method of embodiment 56, wherein the polynucleotide isadministered at an amount between about 0.10 μg per tumor and about 1000mg per tumor.

E58. The method of any one of embodiments 53 to 57, further comprisingadministering an anti-cancer agent.

E59. The method of embodiment 58, wherein the anti-cancer agentcomprises (i) an antibody or antigen-binding fragment thereof thatspecifically binds to PD-1 or PD-L1 (anti-PD-1 antibody or anti-PD-L1antibody, respectively) or a polynucleotide encoding the anti-PD-1 oranti-PD-L1 antibody or antigen-binding fragment thereof, (ii) anantibody or antigen-binding fragment thereof that specifically binds toCTLA-4 (anti-CTLA-4 antibody) or a polynucleotide encoding theanti-CTLA-4 antibody or antigen-binding fragment thereof, or (iii) ananti-PD-1 or anti-PD-L1 antibody or antigen-binding fragment thereof ora polynucleotide encoding the anti-PD-1 or anti-PD-LI antibody orantigen-binding fragment thereof, and an anti-CTLA-4 antibody orantigen-binding fragment thereof or a polynucleotide encoding theanti-CTLA-4 antibody or antigen-binding fragment thereof.

E60. The method of embodiment 58 or 59, wherein the administrationreduces the size of a tumor or inhibits growth of a tumor at least 1.5fold, at least 2 fold, at least 2.5 fold, at least 3 fold, at least 3.5fold, at least 4 fold, at least 4.5 fold, or at least 5 fold better than(i) an administration of a polynucleotide encoding IL12 alone, (ii) anadministration of the anti-PD-1 or anti-PD-L1 antibody alone, or (iii)an administration of the anti-CTLA-4 antibody alone.

E61. The method of embodiment 59 or 60, wherein the polynucleotideencoding the anti-PD-1, anti-PD-L1, or anti-CTLA-4 antibody orantigen-binding fragment thereof comprises an mRNA.

E62. The method of any one of embodiments 59 to 61, wherein thepolynucleotide encoding the anti-PD-1, anti-PD-L1, or anti-CTLA-4antibody or antigen-binding fragment thereof comprises at least onechemically modified nucleoside.

E63. The method of embodiment 62, wherein the at least one chemicallymodified nucleoside is selected from the group consisting of any ofthose listed in Section 13 and a combination thereof.

E64. The method of embodiment 63, wherein the at least one chemicallymodified nucleoside is selected from the group consisting ofpseudouridine, N1-methylpseudouridine, 5-methylcytosine,5-methoxyuridine, and a combination thereof.

E65. The method of any one of embodiments 62 to 64, wherein the mRNAencoding the anti-PD-1, anti-PD-L1, or anti-CTLA-4 antibody orantigen-binding fragment thereof comprises an open reading frame.

E66. The method of any one of embodiments 59 to 65, wherein theanti-PD-L1 antibody is atezolizumab, avelumab, or durvalumab.

E67. The method of any one of embodiments 59 to 66, wherein theanti-CTLA-4 antibody is tremelimumab or ipilimumab.

32. Equivalents and Scope

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments in accordance with the disclosure described herein. Thescope of the present disclosure is not intended to be limited to theabove Description, but rather is as set forth in the appended claims.

In the claims, articles such as “a,” “an,” and “the” may mean one ormore than one unless indicated to the contrary or otherwise evident fromthe context. Claims or descriptions that include “or” between one ormore members of a group are considered satisfied if one, more than one,or all of the group members are present in, employed in, or otherwiserelevant to a given product or process unless indicated to the contraryor otherwise evident from the context. The disclosure includesembodiments in which exactly one member of the group is present in,employed in, or otherwise relevant to a given product or process. Thedisclosure includes embodiments in which more than one, or all of thegroup members are present in, employed in, or otherwise relevant to agiven product or process.

It is also noted that the term “comprising” is intended to be open andpermits but does not require the inclusion of additional elements orsteps. When the term “comprising” is used herein, the term “consistingof” is thus also encompassed and disclosed.

Where ranges are given, endpoints are included. Furthermore, it is to beunderstood that unless otherwise indicated or otherwise evident from thecontext and understanding of one of ordinary skill in the art, valuesthat are expressed as ranges can assume any specific value or subrangewithin the stated ranges in different embodiments of the disclosure, tothe tenth of the unit of the lower limit of the range, unless thecontext clearly dictates otherwise.

In addition, it is to be understood that any particular embodiment ofthe present disclosure that falls within the prior art can be explicitlyexcluded from any one or more of the claims. Since such embodiments aredeemed to be known to one of ordinary skill in the art, they can beexcluded even if the exclusion is not set forth explicitly herein. Anyparticular embodiment of the compositions of the disclosure (e.g., anynucleic acid or protein encoded thereby; any method of production; anymethod of use; etc.) can be excluded from any one or more claims, forany reason, whether or not related to the existence of prior art.

All cited sources, for example, references, publications, databases,database entries, and art cited herein, are incorporated into thisapplication by reference, even if not expressly stated in the citation.In case of conflicting statements of a cited source and the instantapplication, the statement in the instant application shall control.

Section and table headings are not intended to be limiting. The presentdisclosure is further described in the following examples, which do notlimit the scope of the disclosure described in the claims.

EXAMPLES Example 1: In Vitro Expression of Tethered IL-12 mRNA

The in vitro expression of exemplary tethered IL-12 polypeptides encodedby mRNA was assessed. Exemplary schematics of tethered IL-12polypeptides are shown in FIGS. 1A-1D.

A. Preparation of mRNAs

Polynucleotides were prepared comprising nucleotide sequences encoding atethered IL-12 polypeptide (murine IL-12) and a miRNA binding site(miR-122) in the 3′ UTR. The polynucleotide sequences were: mIL12-8TM(SEQ ID NO: 185), encoding a mouse IL-12 polypeptide connected by alinker to a mouse CD8 transmembrane domain (see FIG. 2A); mIL12-PTM (SEQID NO: 183), encoding a mouse IL-12 polypeptide connected by a linker toa mouse PGFRB transmembrane domain (see FIG. 2B); and mIL12-80TM (SEQ IDNO: 181), encoding a mouse IL-12 polypeptide fused to a mouse CD-80transmembrane domain without a linker (see FIG. 2D). The amino acidsequences encoded by the polynucleotides are shown in SEQ ID NOs: 186,184 and 182, respectively. The mRNA open reading frame sequences areshown in SEQ ID NOs: 270, 269 and 268, respectively.

A polynucleotide was also prepared comprising a nucleotide sequenceencoding a secreted IL-12 polypeptide (murine IL-12) and a miRNA bindingsite (miR-122) in its 3′ UTR (mIL12AB; SEQ ID NO: 267).

B. Expression of Tethered IL-12 mRNAs

HeLa cells were seeded on 6-well plates (BD Biosciences, San Jose, USA)one day prior to transfection. Next, mRNAs comprising mIL12AB,mIL12-8TM, mIL12-PTM, or mIL12-80TM mRNA were individually transfectedinto the HeLa cells using 2 μg mRNA and 4 μL Lipofectamine 2000 in 150μL OPTI-MEM per well and incubated. HeLa cells exposed to transfectionreagent with no mRNA served as a negative control (i.e., “Mock”).Transfection media was removed after 4 hours and replaced with freshgrowth medium for the remainder of the incubation period. After 24hours, supernatant was collected from each well, and the cells in eachwell were lysed using a consistent amount of lysis buffer. The amount ofIL-12 (ng/well) in the supernatant and lysate for each well was thenquantified by a standard ELISA assay.

C. Results

FIG. 3 shows that tethered IL-12 containing a linker between the IL-12polypeptide and the transmembrane domain (encoded by mIL12-8TM andmIL12-PTM) was highly expressed in lysate, with low levels detectable inthe supernatant. Tethered mIL-12 in which the transmembrane domain wasfused to the IL-12 polypeptide without a linker (encoded by mIL12-80TM)showed reduced expression in the lysate compared to tethered IL-12containing the linker.

In contrast to tethered IL-12, FIG. 3 shows that secreted mIL-12(encoded by mIL12AB) was highly expressed in supernatant and wasundetectable in the lysate.

Example 2: In Vitro Induction of CD8+ T Cell Proliferation andInterferon-Gamma (IFNγ) Secretion by Tethered IL-12

The in vitro bioactivity of exemplary tethered IL-12 polypeptidesencoded by mRNA was assessed. Specifically, the constructs described inExample 1 were utilized.

A. Preparation of Cultures

CD8+ T cells were isolated from spleens of C57Bl/6 mice using theEasySep™ Mouse CD8+ T Cell Isolation Kit (STEMCELL™ Technologies Inc.,Vancouver, British Columbia, Canada) according to manufacturer protocolsand cultured under standard conditions.

HeLa cell cultures were prepared as described in Example 1, withindividual cultures transfected with mIL12AB, mIL12-8TM, mIL12-PTM, ormIL12-80TM mRNA. HeLa cells exposed to transfection reagent with no mRNAserved as a negative control (“Mock”). HeLa cells were growth arrestedby treating with 50 μg/mL Mitomycin C (Abcam, Cambridge, Mass.) for 20minutes at 37° C., and washed up to four times with growth medium priorto harvesting and seeding for cultures with T cells.

To assess bioactivity, 50,000 CD8+ T cells were cultured with 25,000Dynabeads™ Mouse T-Activator CD3/CD28 beads (ThermoFisher Scientific,Waltham, Mass.) and a fixed number of Mitomycin C-treated HeLa cellsfrom a HeLa cell culture transfected with one of the noted constructsand further including a fixed dilution of supernatant from the HeLa cellculture. Recombinant mouse IL-12 (rmIL12) was also added to a subset ofnegative control cultures (“Mock+rmIL12”). After 72 hours of culture,the proliferation of CD8+ T cells in each culture was determined usingthe CellTiter-Glo® Luminescent Cell Viability Assay kit (PromegaCorporation, Madison, Wis.) according to the manufacturer'sinstructions, and the amount of IFNγ secreted by the same culture wasdetermined by a standard ELISA assay.

B. Results

FIGS. 4A and 4B show that tethered IL-12 containing a linker between theIL-12 polypeptide and the transmembrane domain (encoded by mIL12-8TM andmIL12-PTM) induced higher levels of CD8+ T cell proliferation and IFNγsecretion in the cultures at 72 hours as compared to tethered IL-12lacking a linker between the IL-12 polypeptide and the transmembranedomain (encoded by mIL12-80TM).

A separate assay was performed in which the same cultures were produced(excluding cultures with HeLa cells transfected with mIL12-80TM) andanalyzed for CD8+ T cell proliferation and IFNγ secretion. FIGS. 4C and4D show the results of the assay and demonstrate that tethered IL-12containing a linker between the IL-12 polypeptide and the transmembranedomain (encoded by mIL12-8TM and mIL12-PTM) induced CD8+ T cellproliferation and IFNγ secretion similarly to secreted IL-12 (encoded bymIL12AB).

Example 3: In Vitro Expression and Induction of CD8+ T CellProliferation and Interferon-Gamma (IFNγ) Secretion of Tethered IL-12mRNA

The in vitro expression and bioactivity of exemplary tethered IL-12polypeptides encoded by mRNA was assessed. The constructs described inExample 1, along with additional constructs encoding tethered IL-12polypeptides were utilized.

A. Preparation of mRNAs

Further to the constructs described in Example 1, the followingconstructs were prepared: mIL12-80TID (SEQ ID NO: 236), encoding a mouseIL-12 polypeptide connected by a linker to a mouse CD80 transmembranedomain and intracellular domain (see FIG. 2C); and IgK_mscIL12-80TID(SEQ ID NO: 238), encoding the construct described by Wen-Yu Pan et al.,Mol Therap, Vol. 20(5): 927-937, May 2012 (see FIG. 2E). The amino acidsequences encoded by the polynucleotides are shown in SEQ ID NOs: 237and 239, respectively. The mRNA open reading frame sequences are shownin SEQ ID NOs: 271 and 272, respectively.

B. Expression of Tethered IL-12 mRNAs

HeLa cells were cells seeded on 6-well plates (BD Biosciences, San Jose,USA) one day prior to transfection. Next, mRNAs comprising mIL12AB,mIL12-8TM, mIL12-PTM, mIL12-80TM, mIL12-80TID, or IgK_mscIL12-80TID mRNAwere individually transfected into the HeLa cells using 2 μg mRNA and 4μL Lipofectamine 2000 in 150 μL OPTI-MEM per well and incubated. HeLacells exposed to transfection reagent with no mRNA served as a negativecontrol (i.e.,_“Mock”). Transfection media was removed after 4 hours andreplaced with fresh growth medium for the remainder of the incubationperiod. After 24 hours, supernatant was collected from each well, andthe cells in each well were lysed using a consistent amount of lysisbuffer. The amount of IL-12 (ng/well) in the supernatant and lysate foreach well was then quantified by a standard ELISA assay.

C. Preparation of Cultures

CD8+ T cells were isolated from spleens of C57Bl/6 mice using theEasySep™ Mouse CD8+ T Cell Isolation Kit (STEMCELL™ Technologies Inc.,Vancouver, British Columbia, Canada) according to manufacturer protocolsand cultured under standard conditions.

HeLa cell cultures were prepared as described in Example 1, withindividual cultures transfected with mIL12AB, mIL12-8TM, mIL12-PTM,mIL12-80TM, mIL12-80TID, or IgK_mscIL12-80TID mRNA. HeLa cells exposedto transfection reagent with no mRNA served as a negative control(“Mock”). HeLa cells were growth arrested by treating with 50 pg/mLMitomycin C (Abcam, Cambridge, Mass.) for 20 minutes at 37° C., andwashed up to four times with growth medium prior to harvesting andseeding for cultures with T cells.

To assess bioactivity, 50,000 CD8+ T cells were cultured with 25,000Dynabeads™ Mouse T-Activator CD3/CD28 beads (ThermoFisher Scientific,Waltham, Mass.) and a fixed number of Mitomycin C-treated HeLa cellsfrom a HeLa cell culture transfected with one of the noted constructsand further including a fixed dilution of supernatant from the HeLa cellculture. Recombinant mouse IL-12 (rmIL12) was also added to a subset ofnegative control cultures (“Mock+rmIL12”). After 72 hours of culture,the amount of IFNγ secreted in each culture was determined by a standardELISA assay.

D. Results

FIG. 5A shows expression of IL-12 in the supernatant or lysate, whereinmore IL-12 expression was observed in the lysate in cells transfectedwith mRNA encoding tethered IL-12 polypeptides. FIG. 5B shows inductionof IFNγ secretion by the tethered IL-12 polypeptides was similar toinduction by recombinant IL-12 protein, thereby confirming thebioactivity of the polypeptides encoded by the mRNA constructs. Notably,the constructs encoding tethered IL-12 polypeptides that include alinker between the IL-12 polypeptide and the transmembrane domainexhibited increased protein expression and IFNγ secretion compared toconstructs which lack a linker such as that previously described byWen-Yu Pan et al and mIL12-80TM.

Example 4: In Vivo Effects of Tethered IL-12 mRNA

The in vivo effects of an exemplary tethered IL-12 polypeptide encodedby mRNA was assessed.

A. Preparation of mRNAs

mRNA encoding mIL12-PTM (“tethered mIL-12”) or mIL12 (“secreted mIL-12”)as described in the above Examples was formulated in PEG-DMG lipidnanoparticles (LNP), comprising Compound 18 as the ionizable aminolipid. See U.S. Patent Pub. 2010/0324120, incorporated herein byreference in its entirety. A negative control mRNA (“NST-OX40L”) wasprepared comprising a nucleotide sequence encoding mouse OX40L and amiRNA binding site (miR-122) in the 3′ UTR, along with multiple stopcodons preventing translation of the mRNA (i.e., the sequence encodingOX40L was non-translatable).

B. Mouse Tumor Model

MC38 colon adenocarcinoma tumors were established subcutaneously inC57BL/6 mice by inoculating 5×10⁵ MC38 tumor cells. Once the tumorsreached a mean size of approximately 100 mm³, each animal was treatedwith a single intratumoral dose (5.0 μg/dose) of either mIL12-PTM mRNA(N=20), mIL12 mRNA (N=20), or NST-OX40L (negative control) mRNA (N=20)formulated in the PEG-DMG lipid nanoparticles.

Plasma levels of IL-12 and IFNγ were measured over time using a Luminexbead-based multi-analyte immunoassay (ThermoFisher Scientific, Waltham,Mass.), along with body weight.

C. Results

FIGS. 6A and 6B show the plasma levels of IL-12 and IFNγ, respectively.Tables 2 and 3 below also provide the plasma levels, along with thesecreted:tethered ratio. These results show lower systemic levels ofIL-12 and IFNγ with tethered IL-12 polypeptides, which indicates atolerability benefit.

TABLE 2 Plasma IL-12 Levels Treatment AUC (pg/ml*hr₀₋₁₆₈) Untreated1,090 NST-OX40L 99 Secreted mIL-12 9,496,663 Tethered mIL-12 6,534Secreted:Tethered Ratio 1448x

TABLE 3 Plasma IFNγ Levels Treatment AUC (pg/ml*hr₀₋₁₆₈) Untreated 2,784NST-OX40L 673 Secreted mIL-12 1,853,660 Tethered mIL-12 180,441Secreted:Tethered Ratio 10x

FIG. 7 shows the percentage of body weight change in mice that receivedNST-OX40L (negative control), mIL12 (secreted IL-12) or mIL12-PTM(tethered IL-12). These results indicate tethered IL-12 has less impacton body weight compared to secreted IL-12.

Example 5: In Vivo Anti-Tumor Efficacy of Tethered IL-12 mRNA on Treatedand Distal Tumors in a Dual Tumor Model

The in vivo anti-tumor efficacy of an exemplary tethered IL-12polypeptide encoded by mRNA was assessed.

A. Preparation of mRNAs

mIL12-PTM mRNA was prepared according to Example 4. A negative controlmRNA (“NST-OX40L”) was prepared comprising a nucleotide sequenceencoding mouse OX40L and a miRNA binding site (miR-122) in the 3′ UTR,along with multiple stop codons preventing translation of the mRNA(i.e., the sequence encoding OX40L was non-translatable).

B. Mouse Dual Tumor Model

MC38 colon adenocarcinoma tumors were established subcutaneously inC57BL/6 mice by inoculating 5×10⁵ MC38 tumor cells in both the right(primary) and left (secondary) flanks as shown in FIG. 8A. See alsoRosenberg et al., Science 233(4770):1318-21 (1986).

Once the tumors reached a mean size of approximately 100 mm³, theprimary tumor in the right flank of each animal was treated with eithera single intratumoral dose (5.0 μg/dose) of mIL12-PTM mRNA (N=20) orNST-OX40L (negative control) mRNA (N=20) formulated in PEG-DMG lipidnanoparticles (LNP) prepared according to Example 1. The secondary tumorin the left flank of each animal was not treated.

Tumor volume in the primary tumor and the secondary tumor in each animalwas measured using manual calipers at the indicated time points shown inthe x-axis of FIGS. 8B-8E. Tumor volume was recorded in cubicmillimeters.

C. Results

Intratumoral injection of NST-OX40L negative control mRNA into primarytumors had no effect on tumor volume (see FIG. 8B). Injection of thenegative control mRNA in the primary tumor also had no effect on tumorvolume of untreated secondary tumors in the same animals (see FIG. 8C).

In contrast, intratumoral injection of IL12-PTM mRNA into primary tumorselicited complete responses (no measurable tumor volume) in the primarytumors of 4 animals and partial responses (with tumor volumes of lessthan 60 mm³) in the primary tumors of 2 animals (see FIG. 8D). And, acomplete response (no measurable tumor volume) was observed in theuntreated secondary tumors of 4 animals following injection of IL12-PTMmRNA into the primary tumors of those animals (see FIG. 8E). This resultdemonstrates that treatment with tethered IL-12 mRNA can produce anabscopal effect on an untreated tumor.

Example 6: In Vitro Expression and Induction of CD8+ T CellProliferation and Interferon-Gamma (IFNγ) Secretion of Tethered HumanIL-12 mRNA

The in vitro expression and bioactivity of exemplary tethered humanIL-12 polypeptides encoded by mRNA was assessed.

A. Preparation of mRNAs

Polynucleotides were prepared comprising nucleotide sequences encoding atethered IL-12 polypeptide (human IL-12) and a miRNA binding site(miR-122) in the 3′ UTR. The polynucleotide sequences were: hIL12-8TM(SEQ ID NO: 240), encoding a human IL-12 polypeptide connected by alinker to a human CD8 transmembrane domain, with a V5 tag (see FIG. 9A);hIL12-80TID (SEQ ID NOs: 244-248), encoding a human IL-12 polypeptideconnected by a linker to a human CD80 transmembrane and intracellulardomain (see FIG. 9B); hIL12-PTID570 (SEQ ID NO: 252), encoding a humanIL-12 polypeptide connected by a linker to a human PGFRB transmembranedomain and truncated intracellular domain (E570tr, see amino acidsequence set forth in SEQ ID NO: 227) (see FIG. 9C); and hIL12-PTID739(SEQ ID NO: 254), encoding a human IL-12 polypeptide connected by alinker to a human PGFRB transmembrane domain and truncated intracellulardomain (G739tr, see amino acid sequence set forth in SEQ ID NO: 228)(see FIG. 9D). The amino acid sequences encoded by the polynucleotidesare shown in SEQ ID NOs: 241, 249, 253 and 255, respectively. The mRNAopen reading frame sequences are shown in SEQ ID NOs: 273, 275-279, 281and 282, respectively. Each mRNA comprised a 5′UTR having the sequenceset forth in SEQ ID NO: 287, and 3′UTR having the sequence set forth inSEQ ID NO: 283.

A polynucleotide was also prepared comprising a nucleotide sequenceencoding a secreted IL-12 polypeptide (human IL-12) and a miRNA bindingsite (miR-122) in its 3′ UTR (hIL12AB_041; ORF set forth in SEQ ID NO:221). Table 4 provides correlating amino acid numbering in SEQ ID NO:48, nucleotide numbering in SEQ ID NOs: 5-44, and the 5′ UTR, IL-12Bsignal peptide, mature IL-12A and IL-12B peptides, and linker.

TABLE 4 Amino Acids Nucleotides Signal Peptide IL-12B 1-22 of SEQ ID NO:48 1-66 of SEQ ID NOs: 5-44 Mature IL-12B 23-328 of SEQ ID NO: 48 67-984of SEQ ID NOs: 5-44 Linker 329-335 of SEQ ID NO: 48 985-1005 of SEQ IDNOs: 5-44 Mature IL-12A 336-532 of SEQ ID NO: 48 1006-1596 of SEQ IDNOs: 5-44

B. Expression of Tethered IL-12 mRNAs

HeLa cells were seeded on 6-well plates (BD Biosciences, San Jose, USA)one day prior to transfection. Next, mRNAs comprising hIL12AB_041,hIL12-8TM, hIL12-PTID739, hIL12-PTID570 or hIL12-80TID mRNA wereindividually transfected into the HeLa cells using 2 μg mRNA and 4 μLLipofectamine 2000 in 150 μL OPTI-MEM per well and incubated. HeLa cellsexposed to transfection reagent with no mRNA served as a negativecontrol (i.e., “Mock”). Transfection media was removed after 4 hours andreplaced with fresh growth medium for the remainder of the incubationperiod. After 24 hours, supernatant was collected from each well, andthe cells in each well were lysed using a consistent amount of lysisbuffer. The amount of IL-12 (ng/well) in the supernatant and lysate foreach well was then quantified by a standard ELISA assay.

C. Preparation of Cultures

Peripheral blood mononuclear cells were prepared from whole human bloodby density gradient centrifugation with Lymphoprep™ (STEMCELL™Technologies Inc., Vancouver, British Columbia, Canada) and SepMate™-50tubes (STEMCELL™ Technologies Inc., Vancouver, British Columbia, Canada)according to manufacturer protocols. CD8+ T cells were then isolatedusing the EasySep™ Human CD8+ T Cell Isolation Kit (STEMCELL™Technologies Inc., Vancouver, British Columbia, Canada) according tomanufacturer protocols.

HeLa cell cultures were prepared as described in Example 1, withindividual cultures transfected with hIL12AB_002, hIL12-8TM,hIL12-PTID739, hIL12-PTID570 or hIL12-80TID mRNA. HeLa cells exposed totransfection reagent with no mRNA served as a negative control (“Mock”).HeLa cells were growth arrested by treating with 50 μg/mL Mitomycin C(Abcam, Cambridge, Mass.) for 20 minutes at 37° C., and washed up tofour times with growth medium prior to harvesting and seeding forcultures with T cells.

To assess bioactivity, 75,000 human peripheral blood CD8+ T cells werecultured with 25,000 Dynabeads™ Human T-Activator CD3/CD28 beads(ThermoFisher Scientific, Waltham, Mass.) and a fixed number ofMitomycin C-treated HeLa cells from a HeLa cell culture transfected withone of the noted constructs and further including a fixed dilution ofsupernatant from the HeLa cell culture. Recombinant human IL-12 (rhIL12)was also added to a subset of negative control cultures (“Mock+rhIL12”).After 48 hours of culture, the amount of IFNγ secreted in each culturewas determined by a standard ELISA assay.

D. Results

FIG. 10A shows expression of IL-12 in the supernatant or lysate, whereinmore IL-12 expression was observed in the lysate in cells transfectedwith mRNA encoding tethered IL-12 polypeptides. FIG. 10B shows inductionof IFNγ secretion by the tethered IL-12 polypeptides was similar toinduction by recombinant IL-12 protein, thereby confirming thebioactivity of the polypeptides encoded by the mRNA constructs. Tetheredconstructs demonstrate comparable bioactivity as secreted constructs,while demonstrating almost undetectable IL-12 expression in supernatant.This result suggests that these constructs should also have far lowersystemic exposure in vivo than a secreted IL-12, which could provide atolerability benefit.

Further, FIG. 11 shows expression of hIL12-80TID encoded by fourdifferent mRNA sequences (SEQ ID NOs: 276, 277, 278 and 279). Theseresults indicated comparable IL-12 expression in the lysate with minimalexpression in the supernatant, regardless of the mRNA used to encode thesame amino acid sequence.

OTHER EMBODIMENTS

It is to be understood that the words which have been used are words ofdescription rather than limitation, and that changes can be made withinthe purview of the appended claims without departing from the true scopeand spirit of the disclosure in its broader aspects.

While the present disclosure has been described at some length and withsome particularity with respect to the several described embodiments, itis not intended that it should be limited to any such particulars orembodiments or any particular embodiment, but it is to be construed withreferences to the appended claims so as to provide the broadest possibleinterpretation of such claims in view of the prior art and, therefore,to effectively encompass the intended scope of the disclosure.

All publications, patent applications, patents, and other referencesmentioned herein are incorporated by reference in their entirety. Incase of conflict, the present specification, including definitions, willcontrol. In addition, section headings, the materials, methods, andexamples are illustrative only and not intended to be limiting.

It is to be appreciated that the Detailed Description section, and notthe Summary and Abstract sections, is intended to be used to interpretthe claims. The Summary and Abstract sections may set forth one or morebut not all exemplary embodiments of the present disclosure ascontemplated by the inventor(s), and thus, are not intended to limit thepresent disclosure and the appended claims in any way.

The present disclosure has been described above with the aid offunctional building blocks illustrating the implementation of specifiedfunctions and relationships thereof. The boundaries of these functionalbuilding blocks have been arbitrarily defined herein for the convenienceof the description. Alternate boundaries can be defined so long as thespecified functions and relationships thereof are appropriatelyperformed.

The foregoing description of the specific embodiments will so fullyreveal the general nature of the disclosure that others can, by applyingknowledge within the skill of the art, readily modify and/or adapt forvarious applications such specific embodiments, without undueexperimentation, without departing from the general concept of thepresent disclosure. Therefore, such adaptations and modifications areintended to be within the meaning and range of equivalents of thedisclosed embodiments, based on the teaching and guidance presentedherein. It is to be understood that the phraseology or terminologyherein is for the purpose of description and not of limitation, suchthat the terminology or phraseology of the present specification is tobe interpreted by the skilled artisan in light of the teachings andguidance.

The breadth and scope of the present disclosure should not be limited byany of the above-described exemplary embodiments, but should be definedonly in accordance with the following claims and their equivalents.

Summary of Sequences

SEQ ID NO Description Sequence   1 Wild TypeIWELKKDVYWELDWYPDAPGEMWLTCDTPEEDGITWTLDQSSEVLGSGKTLTIQ IL12B withoutVKEFGDAGQYTCHKGGEVLSHSLLLLHKKEDGIWSTDILKDQKEPKNKTFLRCEAK signal (IL12B)NYSGRFTCWWLTTISTDLTFSVKSSRGSSDPQGVTCGAATLSAERVRGDNKEYEYS Amino AcidsVECQEDSACPAAEESLPIEVMVDAVHKLKYENYTSSFFIRDIIKPDPPKNLQLKPLKNSRQVEVSWEYPDTWSTPHSYFSLTFCVQVQGKSKREKKDRVFTDKTSATVICRKNASISVRAQDRYYSSSWSEWASVPCS   2 Wild TypeATATGGGAACTGAAGAAAGATGTTTATGTCGTAGAATTGGATTGGTATCCGGATGC IL12B withoutCCCTGGAGAAATGGTGGTCCTCACCTGTGACACCCCTGAAGAAGATGGTATCACCT signal (IL12B)GGACCTTGGACCAGAGCAGTGAGGTCTTAGGCTCTGGCAAAACCCTGACCATCCAA Nucleic AcidsGTCAAAGAGTTTGGAGATGCTGGCCAGTACACCTGTCACAAAGGAGGCGAGGTTCTAAGCCATTCGCTCCTGCTGCTTCACAAAAAGGAAGATGGAATTTGGTCCACTGATATTTTAAAGGACCAGAAAGAACCCAAAAATAAGACCTTTCTAAGATGCGAGGCCAAGAATTATTCTGGACGTTTCACCTGCTGGTGGCTGACGACAATCAGTACTGATTTGACATTCAGTGTCAAAAGCAGCAGAGGCTCTTCTGACCCCCAAGGGGTGACGTGCGGAGCTGCTACACTCTCTGCAGAGAGAGTCAGAGGGGACAACAAGGAGTATGAGTACTCAGTGGAGTGCCAGGAGGACAGTGCCTGCCCAGCTGCTGAGGAGAGTCTGCCCATTGAGGTCATGGTGGATGCCGTTCACAAGCTCAAGTATGAAAACTACACCAGCAGCTTCTTCATCAGGGACATCATCAAACCTGACCCACCCAAGAACTTGCAGCTGAAGCCATTAAAGAATTCTCGGCAGGTGGAGGTCAGCTGGGAGTACCCTGACACCTGGAGTACTCCACATTCCTACTTCTCCCTGACATTCTGCGTTCAGGTCCAGGGCAAGAGCAAGAGAGAAAAGAAAGATAGAGTCTTCACGGACAAGACCTCAGCCACGGTCATCTGCCGCAAAAATGCCAGCATTAGCGTGCGGGCCCAGGACCGCTACTATAGCTCATCTTGGAGCGAATGGGCATCTGTGCCCTGCAGT   3 Wild TypeRNLPVATPDPGMFPCLHHSQNLLRAVSNMLQKARQTLEFYPCTSEEIDHEDITKDK IL12 A withoutTSTVEACLPLELTKNESCLNSRETSFITNGSCLASRKTSFMMALCLSSIYEDLKMY signal peptideQVEFKTMNAKLLMDPKRQIFLDQNMLAVIDELMQALNFNSETVPQKSSLEEPDFYK Amino acidsTKIKLCILLHAFRIRAVTIDRVMSYLNAS   4 Wild TypeAGAAACCTCCCCGTGGCCACTCCAGACCCAGGAATGTTCCCATGCCTTCACCACTC IL12A withoutCCAAAACCTGCTGAGGGCCGTCAGCAACATGCTCCAGAAGGCCAGACAAACTCTAG signal peptideAATTTTACCCTTGCACTTCTGAAGAGATTGATCATGAAGATATCACAAAAGATAAA Nucleic acidsACCAGCACAGTGGAGGCCTGTTTACCATTGGAATTAACCAAGAATGAGAGTTGCCTAAATTCCAGAGAGACCTCTTTCATAACTAATGGGAGTTGCCTGGCCTCCAGAAAGACCTCTTTTATGATGGCCCTGTGCCTTAGTAGTATTTATGAAGACTTGAAGATGTACCAGGTGGAGTTCAAGACCATGAATGCAAAGCTTCTGATGGATCCTAAGAGGCAGATCTTTCTAGATCAAAACATGCTGGCAGTTATTGATGAGCTGATGCAGGCCCTGAATTTCAACAGTGAGACTGTGCCACAAAAATCCTCCCTTGAAGAACCGGATTTTTATAAAACTAAAATCAAGCTCTGCATACTTCTTCATGCTTTCAGAATTCGGGCAGTGACTATTGATAGAGTGATGAGCTATCTGAATGCTTCC   5 hIL12AB_001ATGTGTCACCAGCAGCTGGTCATTAGCTGGTTTAGCCTTGTGTTCCTGGCCTCCCC ORFCCTTGTCGCTATTTGGGAGCTCAAGAAGGACGTGTACGTGGTGGAGCTGGACTGGTACCCAGACGCGCCCGGAGAGATGGTAGTTCTGACCTGTGATACCCCAGAGGAGGACGGCATCACCTGGACTCTGGACCAAAGCAGCGAGGTTTTGGGCTCAGGGAAAACGCTGACCATCCAGGTGAAGGAATTCGGCGACGCCGGACAGTACACCTGCCATAAGGGAGGAGAGGTGCTGAGCCATTCCCTTCTTCTGCTGCACAAGAAAGAGGACGGCATCTGGTCTACCGACATCCTGAAAGACCAGAAGGAGCCCAAGAACAAAACCTTCCTGAGGTGCGAGGCCAAGAACTACTCCGGCAGGTTCACTTGTTGGTGGCTGACCACCATCAGTACAGACCTGACTTTTAGTGTAAAAAGCTCCAGAGGCTCGTCCGATCCCCAAGGGGTGACCTGCGGCGCAGCCACTCTGAGCGCTGAGCGCGTGCGCGGTGACAATAAAGAGTACGAGTACAGCGTTGAGTGTCAAGAAGACAGCGCTTGCCCTGCCGCCGAGGAGAGCCTGCCTATCGAGGTGATGGTTGACGCAGTGCACAAGCTTAAGTACGAGAATTACACCAGCTCATTCTTCATTAGAGATATAATCAAGCCTGACCCACCCAAGAACCTGCAGCTGAAGCCACTGAAAAACTCACGGCAGGTCGAAGTGAGCTGGGAGTACCCCGACACCTGGAGCACTCCTCATTCCTATTTCTCTCTTACATTCTGCGTCCAGGTGCAGGGCAAGAGCAAGCGGGAAAAGAAGGATCGAGTCTTCACCGACAAAACAAGCGCGACCGTGATTTGCAGGAAGAACGCCAGCATCTCCGTCAGAGCCCAGGATAGATACTATAGTAGCAGCTGGAGCGAGTGGGCAAGCGTGCCCTGTTCCGGCGGCGGGGGCGGGGGCAGCCGAAACTTGCCTGTCGCTACCCCGGACCCTGGAATGTTTCCGTGTCTGCACCACAGCCAGAACCTGCTGAGAGCCGTGTCGAATATGCTCCAGAAGGCCCGGCAGACCCTTGAGTTCTACCCCTGTACCAGCGAAGAGATCGATCATGAGGACATCACGAAAGACAAGACTTCCACCGTCGAGGCTTGTCTCCCGCTGGAGCTGACCAAGAACGAGAGCTGTCTGAATAGCCGGGAGACATCTTTCATCACGAATGGTAGCTGTCTGGCCAGCAGGAAAACTTCCTTCATGATGGCTCTCTGCCTGAGCTCTATCTATGAAGATCTGAAGATGTATCAGGTGGAGTTTAAGACTATGAACGCCAAACTCCTGATGGACCCAAAAAGGCAAATCTTTCTGGACCAGAATATGCTGGCCGTGATAGACGAGCTGATGCAGGCACTGAACTTCAACAGCGAGACAGTGCCACAGAAATCCAGCCTGGAGGAGCCTGACTTTTACAAAACTAAGATCAAGCTGTGTATCCTGCTGCACGCCTTTAGAATCCGTGCCGTGACTATCGACAGGGTGATGTCATACCTCAACGCTTCA   6 hIL12AB_002ATGTGCCACCAGCAGCTGGTGATCAGCTGGTTCAGCCTGGTGTTCCTGGCCAGCCC ORFCCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGCTGGACTGGTACCCCGACGCCCCCGGCGAGATGGTGGTGCTGACCTGCGACACCCCCGAGGAGGACGGCATCACCTGGACCCTGGACCAGAGCAGCGAGGTGCTGGGCAGCGGCAAGACCCTGACCATCCAGGTGAAGGAGTTCGGCGACGCCGGCCAGTACACCTGCCACAAGGGCGGCGAGGTGCTGAGCCACAGCCTGCTGCTGCTGCACAAGAAGGAGGACGGCATCTGGAGCACCGACATCCTGAAGGACCAGAAGGAGCCCAAGAACAAGACCTTCCTGAGATGCGAGGCCAAGAACTACAGCGGCAGATTCACCTGCTGGTGGCTGACCACCATCAGCACCGACCTGACCTTCAGCGTGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGTGACCTGCGGCGCCGCCACCCTGAGCGCCGAGAGAGTGAGAGGCGACAACAAGGAGTACGAGTACAGCGTGGAGTGCCAGGAGGACAGCGCCTGCCCCGCCGCCGAGGAGAGCCTGCCCATCGAGGTGATGGTGGACGCCGTGCACAAGCTGAAGTACGAGAACTACACCAGCAGCTTCTTCATCAGAGACATCATCAAGCCCGACCCCCCCAAGAACCTGCAGCTGAAGCCCCTGAAGAACAGCAGACAGGTGGAGGTGAGCTGGGAGTACCCCGACACCTGGAGCACCCCCCACAGCTACTTCAGCCTGACCTTCTGCGTGCAGGTGCAGGGCAAGAGCAAGAGAGAGAAGAAGGACAGAGTGTTCACCGACAAGACCAGCGCCACCGTGATCTGCAGAAAGAACGCCAGCATCAGCGTGAGAGCCCAGGACAGATACTACAGCAGCAGCTGGAGCGAGTGGGCCAGCGTGCCCTGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCTGCCCGTGGCCACCCCCGACCCCGGCATGTTCCCCTGCCTGCACCACAGCCAGAACCTGCTGAGAGCCGTGAGCAACATGCTGCAGAAGGCCAGACAGACCCTGGAGTTCTACCCCTGCACCAGCGAGGAGATCGACCACGAGGACATCACCAAGGACAAGACCAGCACCGTGGAGGCCTGCCTGCCCCTGGAGCTGACCAAGAACGAGAGCTGCCTGAACAGCAGAGAGACCAGCTTCATCACCAACGGCAGCTGCCTGGCCAGCAGAAAGACCAGCTTCATGATGGCCCTGTGCCTGAGCAGCATCTACGAGGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAACGCCAAGCTGCTGATGGACCCCAAGAGACAGATCTTCCTGGACCAGAACATGCTGGCCGTGATCGACGAGCTGATGCAGGCCCTGAACTTCAACAGCGAGACCGTGCCCCAGAAGAGCAGCCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATCCTGCTGCACGCCTTCAGAATCAGAGCCGTGACCATCGACAGAGTGATGAGCTACCTGAACGCCAGC   7 hIL12AB_003ATGTGTCACCAGCAGTTGGTCATCTCTTGGTTTTCCCTGGTTTTTCTGGCATCTCC ORFCCTCGTGGCCATATGGGAACTGAAGAAAGATGTTTATGTCGTAGAATTGGATTGGTATCCGGATGCCCCTGGAGAAATGGTGGTCCTCACCTGTGACACCCCTGAAGAAGATGGTATCACCTGGACCTTGGACCAGAGCAGTGAGGTCTTAGGCTCTGGCAAAACCCTGACCATCCAAGTCAAAGAGTTTGGAGATGCTGGCCAGTACACCTGTCACAAAGGAGGCGAGGTTCTAAGCCATTCGCTCCTGCTGCTTCACAAAAAGGAAGATGGAATTTGGTCCACTGATATTTTAAAGGACCAGAAAGAACCCAAAAATAAGACCTTTCTAAGATGCGAGGCCAAGAATTATTCTGGACGTTTCACCTGCTGGTGGCTGACGACAATCAGTACTGATTTGACATTCAGTGTCAAAAGCAGCAGAGGCTCTTCTGACCCCCAAGGGGTGACGTGCGGAGCTGCTACACTCTCTGCAGAGAGAGTCAGAGGGGACAACAAGGAGTATGAGTACTCAGTGGAGTGCCAGGAGGACAGTGCCTGCCCAGCTGCTGAGGAGAGTCTGCCCATTGAGGTCATGGTGGATGCCGTTCACAAGCTCAAGTATGAAAACTACACCAGCAGCTTCTTCATCAGGGACATCATCAAACCTGACCCACCCAAGAACTTGCAGCTGAAGCCATTAAAGAATTCTCGGCAGGTGGAGGTCAGCTGGGAGTACCCTGACACCTGGAGTACTCCACATTCCTACTTCTCCCTGACATTCTGCGTTCAGGTCCAGGGCAAGAGCAAGAGAGAAAAGAAAGATAGAGTCTTCACGGACAAGACCTCAGCCACGGTCATCTGCCGCAAAAATGCCAGCATTAGCGTGCGGGCCCAGGACCGCTACTATAGCTCATCTTGGAGCGAATGGGCATCTGTGCCCTGCAGTGGCGGAGGGGGCGGAGGGAGCAGAAACCTCCCCGTGGCCACTCCAGACCCAGGAATGTTCCCATGCCTTCACCACTCCCAAAACCTGCTGAGGGCCGTCAGCAACATGCTCCAGAAGGCCAGACAAACTTTAGAATTTTACCCTTGCACTTCTGAAGAGATTGATCATGAAGATATCACAAAAGATAAAACCAGCACAGTGGAGGCCTGTTTACCATTGGAATTAACCAAGAATGAGAGTTGCCTAAATTCCAGAGAGACCTCTTTCATAACTAATGGGAGTTGCCTGGCCTCCAGAAAGACCTCTTTTATGATGGCCCTGTGCCTTAGTAGTATTTATGAAGACTTGAAGATGTACCAGGTGGAGTTCAAGACCATGAATGCAAAGCTTCTGATGGATCCTAAGAGGCAGATCTTTTTAGATCAAAACATGCTGGCAGTTATTGATGAGCTGATGCAGGCCCTGAATTTCAACAGTGAGACTGTGCCACAAAAATCCTCCCTTGAAGAACCGGACTTCTACAAGACCAAGATCAAGCTCTGCATACTTCTTCATGCTTTCAGAATTCGGGCAGTGACTATTGATAGAGTGATGAGCTATCTGAATGCTTCC   8 hIL12AB_004ATGGGCTGCCACCAGCAGCTGGTCATCAGCTGGTTCTCCCTGGTCTTCCTGGCCAG ORFCCCCCTGGTGGCCATCTGGGAGCTGAAGAAAGATGTCTATGTTGTAGAGCTGGACTGGTACCCAGATGCTCCTGGAGAAATGGTGGTTCTCACCTGTGACACGCCAGAAGAAGATGGCATCACCTGGACGCTGGACCAGAGCTCAGAAGTTCTTGGCAGTGGAAAAACGCTGACCATACAAGTAAAAGAATTTGGGGATGCTGGCCAGTACACCTGCCACAAAGGAGGAGAAGTTCTCAGCCACAGCCTGCTGCTGCTGCACAAGAAAGAAGATGGCATCTGGAGCACAGACATTTTAAAAGACCAGAAGGAGCCCAAGAACAAAACCTTCCTTCGATGTGAGGCCAAGAACTACAGTGGCCGCTTCACCTGCTGGTGGCTCACCACCATCAGCACAGACCTCACCTTCTCGGTGAAGAGCAGCCGTGGCAGCTCAGACCCCCAAGGAGTCACCTGTGGGGCGGCCACGCTGTCGGCAGAAAGAGTTCGAGGGGACAACAAGGAATATGAATACTCGGTGGAATGTCAAGAAGACTCGGCCTGCCCGGCGGCAGAAGAAAGTCTTCCCATAGAAGTCATGGTGGATGCTGTTCACAAATTAAAATATGAAAACTACACCAGCAGCTTCTTCATCAGAGACATCATCAAGCCAGACCCGCCCAAGAACCTGCAGCTGAAGCCCCTGAAGAACAGCAGACAAGTGGAAGTTTCCTGGGAGTACCCAGACACGTGGAGCACGCCGCACAGCTACTTCAGCCTCACCTTCTGTGTACAAGTACAAGGCAAGAGCAAGAGAGAGAAGAAAGATCGTGTCTTCACAGACAAAACCTCGGCGACGGTCATCTGCAGGAAGAATGCCTCCATCTCGGTTCGAGCCCAGGACCGCTACTACAGCAGCAGCTGGAGTGAGTGGGCCTCGGTGCCCTGCAGTGGTGGCGGCGGCGGCGGCAGCAGAAACCTTCCTGTGGCCACGCCGGACCCTGGCATGTTCCCGTGCCTGCACCACAGCCAAAATTTACTTCGAGCTGTTTCTAACATGCTGCAGAAAGCAAGACAAACTTTAGAATTCTACCCCTGCACCTCAGAAGAAATAGACCATGAAGACATCACCAAAGATAAAACCAGCACTGTAGAGGCCTGCCTGCCCCTGGAGCTCACCAAGAATGAATCCTGCCTCAACAGCAGAGAGACCAGCTTCATCACCAATGGCAGCTGCCTGGCCAGCAGGAAAACCAGCTTCATGATGGCGCTCTGCCTGAGCAGCATCTATGAAGATTTGAAGATGTACCAAGTAGAATTTAAAACCATGAATGCCAAGCTGCTCATGGACCCCAAGAGACAAATATTTTTGGATCAAAACATGCTGGCTGTCATTGATGAGCTCATGCAAGCATTAAACTTCAACTCAGAGACGGTGCCCCAGAAGAGCAGCCTGGAGGAGCCAGACTTCTACAAAACCAAGATCAAGCTCTGCATCTTATTACATGCCTTCCGCATCCGGGCGGTCACCATTGACCGTGTCATGTCCTACTTAAATGCCAGC   9 hIL12AB_005ATGTGCCACCAGCAGCTGGTCATCAGCTGGTTCTCCCTGGTCTTCCTGGCCAGCCC ORFCCTGGTGGCCATCTGGGAGCTGAAGAAAGATGTCTATGTTGTAGAGCTGGACTGGTACCCAGATGCTCCTGGAGAAATGGTGGTTCTCACCTGTGACACGCCAGAAGAAGATGGCATCACCTGGACGCTGGACCAGAGCTCAGAAGTTCTTGGCAGTGGAAAAACGCTGACCATACAAGTAAAAGAATTTGGGGATGCTGGCCAGTACACCTGCCACAAAGGAGGAGAAGTTCTCAGCCACAGCCTGCTGCTGCTGCACAAGAAAGAAGATGGCATCTGGAGCACAGACATTTTAAAAGACCAGAAGGAGCCCAAGAACAAAACCTTCCTTCGATGTGAGGCCAAGAACTACAGTGGCCGCTTCACCTGCTGGTGGCTCACCACCATCAGCACAGACCTCACCTTCTCGGTGAAGAGCAGCCGTGGCAGCTCAGACCCCCAAGGAGTCACCTGTGGGGCGGCCACGCTGTCGGCAGAAAGAGTTCGAGGGGACAACAAGGAATATGAATACTCGGTGGAATGTCAAGAAGACTCGGCCTGCCCGGCGGCAGAAGAAAGTCTTCCCATAGAAGTCATGGTGGATGCTGTTCACAAATTAAAATATGAAAACTACACCAGCAGCTTCTTCATCAGAGACATCATCAAGCCAGACCCGCCCAAGAACCTGCAGCTGAAGCCCCTGAAGAACAGCAGACAAGTGGAAGTTTCCTGGGAGTACCCAGACACGTGGAGCACGCCGCACAGCTACTTCAGCCTCACCTTCTGTGTACAAGTACAAGGCAAGAGCAAGAGAGAGAAGAAAGATCGTGTCTTCACAGACAAAACCTCGGCGACGGTCATCTGCAGGAAGAATGCCTCCATCTCGGTTCGAGCCCAGGACCGCTACTACAGCAGCAGCTGGAGTGAGTGGGCCTCGGTGCCCTGCAGTGGTGGCGGCGGCGGCGGCAGCAGAAACCTTCCTGTGGCCACGCCGGACCCTGGCATGTTCCCGTGCCTGCACCACAGCCAAAATTTACTTCGAGCTGTTTCTAACATGCTGCAGAAAGCAAGACAAACTTTAGAATTCTACCCCTGCACCTCAGAAGAAATAGACCATGAAGACATCACCAAAGATAAAACCAGCACTGTAGAGGCCTGCCTGCCCCTGGAGCTCACCAAGAATGAATCCTGCCTCAACAGCAGAGAGACCAGCTTCATCACCAATGGCAGCTGCCTGGCCAGCAGGAAAACCAGCTTCATGATGGCGCTCTGCCTGAGCAGCATCTATGAAGATTTGAAGATGTACCAAGTAGAATTTAAAACCATGAATGCCAAGCTGCTCATGGACCCCAAGAGACAAATATTTTTGGATCAAAACATGCTGGCTGTCATTGATGAGCTCATGCAAGCATTAAACTTCAACTCAGAGACGGTGCCCCAGAAGAGCAGCCTGGAGGAGCCAGACTTCTACAAAACCAAGATCAAGCTCTGCATCTTATTACATGCCTTCCGCATCCGGGCGGTCACCATTGACCGTGTCATGTCCTACTTAAATGCCAGC  10 hIL12AB_006ATGTGCCACCAGCAGCTGGTGATCAGCTGGTTCAGCCTGGTGTTCCTGGCCAGCCC ORFCCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGCTGGACTGGTACCCCGACGCCCCCGGCGAGATGGTGGTGCTGACCTGTGACACCCCCGAGGAGGACGGCATCACCTGGACCCTGGACCAGAGCAGCGAGGTGCTGGGCAGCGGCAAGACCCTGACCATCCAGGTGAAGGAGTTCGGGGACGCCGGCCAGTACACCTGCCACAAGGGCGGCGAGGTGCTGAGCCACAGCCTGCTGCTGCTGCACAAGAAGGAGGACGGCATCTGGAGCACAGATATCCTGAAGGACCAGAAGGAGCCCAAGAACAAGACCTTCCTGAGATGCGAGGCCAAGAACTACAGCGGCAGATTCACCTGCTGGTGGCTGACCACCATCAGCACAGACTTGACCTTCAGCGTGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGTGACCTGCGGCGCCGCCACCCTGAGCGCCGAGAGAGTGAGAGGGGACAACAAGGAGTACGAGTACAGCGTGGAGTGCCAGGAGGACAGCGCCTGCCCCGCCGCCGAGGAGAGCCTGCCCATCGAGGTGATGGTGGACGCCGTGCACAAGCTGAAGTACGAGAACTACACCAGCAGCTTCTTCATCAGAGACATCATCAAGCCCGACCCGCCGAAGAACCTGCAGCTGAAGCCCCTGAAGAACAGCAGACAGGTGGAGGTGAGCTGGGAGTACCCCGACACCTGGAGCACCCCCCACAGCTACTTCAGCCTGACCTTCTGCGTGCAGGTGCAGGGCAAGAGCAAGAGAGAGAAGAAGGACAGAGTGTTCACAGATAAGACCAGCGCCACCGTGATCTGCAGAAAGAACGCCAGCATCAGCGTGAGAGCCCAGGACAGATACTACAGCAGCAGCTGGAGCGAGTGGGCCAGCGTGCCCTGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCTGCCCGTGGCCACCCCCGACCCCGGCATGTTCCCCTGCCTGCACCACAGCCAGAACCTGCTGAGAGCCGTGAGCAACATGCTGCAGAAGGCCAGACAGACCCTGGAGTTCTACCCCTGCACCAGCGAGGAGATCGACCACGAGGACATCACCAAGGACAAGACCAGCACCGTGGAGGCCTGCCTGCCCCTGGAGCTGACCAAGAATGAAAGCTGCCTGAACAGCAGAGAGACCAGCTTCATCACCAACGGCAGCTGCCTGGCCAGCAGAAAGACCAGCTTCATGATGGCCCTGTGCCTGAGCAGCATCTACGAGGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAACGCCAAGCTGCTGATGGACCCCAAGAGACAGATCTTCCTGGACCAGAACATGCTGGCCGTGATCGACGAGCTGATGCAGGCCCTGAACTTCAACAGCGAGACCGTGCCCCAGAAGAGCAGCCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATCCTGCTGCACGCCTTCAGAATCAGAGCCGTGACCATCGACAGAGTGATGAGCTACCTGAACGCCAGC  11 hIL12AB_007ATGTGCCACCAGCAGCTTGTCATCTCCTGGTTCTCTCTTGTCTTCCTTGCTTCTCC ORFTCTTGTGGCCATCTGGGAGCTGAAGAAGGATGTTTATGTTGTGGAGTTGGACTGGTACCCTGATGCTCCTGGAGAAATGGTGGTTCTCACCTGTGACACTCCTGAGGAGGATGGCATCACCTGGACTTTGGACCAGTCTTCTGAGGTTCTTGGCAGTGGAAAAACTCTTACTATTCAGGTGAAGGAGTTTGGAGATGCTGGCCAGTACACCTGCCACAAGGGTGGTGAAGTTCTCAGCCACAGTTTACTTCTTCTTCACAAGAAGGAGGATGGCATCTGGTCTACTGACATTTTAAAAGACCAGAAGGAGCCCAAGAACAAGACTTTCCTTCGTTGTGAAGCCAAGAACTACAGTGGTCGTTTCACCTGCTGGTGGCTTACTACTATTTCTACTGACCTTACTTTCTCTGTGAAGTCTTCTCGTGGCTCTTCTGACCCTCAGGGTGTCACCTGTGGGGCTGCTACTCTTTCTGCTGAGCGTGTGCGTGGGGACAACAAGGAGTATGAATACTCGGTGGAGTGCCAGGAGGACTCTGCCTGCCCTGCTGCTGAGGAGTCTCTTCCTATTGAGGTGATGGTGGATGCTGTGCACAAGTTAAAATATGAAAACTACACTTCTTCTTTCTTCATTCGTGACATTATAAAACCTGACCCTCCCAAGAACCTTCAGTTAAAACCTTTAAAAAACTCTCGTCAGGTGGAGGTGTCCTGGGAGTACCCTGACACGTGGTCTACTCCTCACTCCTACTTCTCTCTTACTTTCTGTGTCCAGGTGCAGGGCAAGTCCAAGCGTGAGAAGAAGGACCGTGTCTTCACTGACAAGACTTCTGCTACTGTCATCTGCAGGAAGAATGCATCCATCTCTGTGCGTGCTCAGGACCGTTACTACAGCTCTTCCTGGTCTGAGTGGGCTTCTGTGCCCTGCTCTGGCGGCGGCGGCGGCGGCAGCAGAAATCTTCCTGTGGCTACTCCTGACCCTGGCATGTTCCCCTGCCTTCACCACTCGCAGAACCTTCTTCGTGCTGTGAGCAACATGCTTCAGAAGGCTCGTCAGACTTTAGAATTCTACCCCTGCACTTCTGAGGAGATTGACCATGAAGACATCACCAAGGACAAGACTTCTACTGTGGAGGCCTGCCTTCCTTTAGAGCTGACCAAGAATGAATCCTGCTTAAATTCTCGTGAGACTTCTTTCATCACCAATGGCAGCTGCCTTGCCTCGCGCAAGACTTCTTTCATGATGGCTCTTTGCCTTTCTTCCATCTATGAAGACTTAAAAATGTACCAGGTGGAGTTCAAGACCATGAATGCAAAGCTTCTCATGGACCCCAAGCGTCAGATATTTTTGGACCAGAACATGCTTGCTGTCATTGATGAGCTCATGCAGGCTTTAAACTTCAACTCTGAGACTGTGCCTCAGAAGTCTTCTTTAGAAGAGCCTGACTTCTACAAGACCAAGATAAAACTTTGCATTCTTCTTCATGCTTTCCGCATCCGTGCTGTGACTATTGACCGTGTGATGTCCTACTTAAATGCTTCT  12 hIL12AB_008ATGTGTCATCAACAACTCGTGATTAGCTGGTTCAGTCTCGTGTTCCTGGCCTCTCC ORFGCTGGTGGCCATCTGGGAGCTTAAGAAGGACGTGTACGTGGTGGAGCTCGATTGGTACCCCGATGCTCCTGGCGAGATGGTGGTGCTAACCTGCGATACCCCCGAGGAGGACGGGATCACTTGGACCCTGGATCAGAGTAGCGAAGTCCTGGGCTCTGGCAAGACACTCACAATCCAGGTGAAGGAATTCGGAGACGCTGGTCAGTACACTTGCCACAAGGGGGGTGAAGTGCTGTCTCACAGCCTGCTGTTACTGCACAAGAAGGAGGATGGGATCTGGTCAACCGACATCCTGAAGGATCAGAAGGAGCCTAAGAACAAGACCTTTCTGAGGTGTGAAGCTAAGAACTATTCCGGAAGATTCACTTGCTGGTGGTTGACCACAATCAGCACTGACCTGACCTTTTCCGTGAAGTCCAGCAGAGGAAGCAGCGATCCTCAGGGCGTAACGTGCGGCGCGGCTACCCTGTCAGCTGAGCGGGTTAGAGGCGACAACAAAGAGTATGAGTACTCCGTGGAGTGTCAGGAGGACAGCGCCTGCCCCGCAGCCGAGGAGAGTCTGCCCATCGAGGTGATGGTGGACGCTGTCCATAAGTTAAAATACGAAAATTACACAAGTTCCTTTTTCATCCGCGATATTATCAAACCCGATCCCCCCAAGAACCTGCAGCTGAAGCCCCTGAAGAATAGCCGACAGGTGGAAGTCTCTTGGGAGTATCCTGACACCTGGTCCACGCCTCACAGCTACTTTAGTCTGACTTTCTGTGTCCAGGTCCAGGGCAAGAGCAAGAGAGAGAAAAAGGATAGAGTGTTTACTGACAAGACATCTGCTACAGTCATCTGCAGAAAGAACGCCAGTATCTCAGTGAGGGCGCAGGACAGATACTACAGTAGTAGCTGGAGCGAATGGGCTAGCGTGCCCTGTTCAGGGGGCGGCGGAGGGGGCTCCAGGAATCTGCCCGTGGCCACCCCCGACCCTGGGATGTTCCCTTGCCTCCATCACTCACAGAACCTGCTCAGAGCAGTGAGCAACATGCTCCAAAAGGCCCGCCAGACCCTGGAGTTTTACCCTTGTACTTCAGAAGAGATCGATCACGAAGACATAACAAAGGATAAAACCAGCACCGTGGAGGCCTGTCTGCCTCTAGAACTCACAAAGAATGAAAGCTGTCTGAATTCCAGGGAAACCTCCTTCATTACTAACGGAAGCTGTCTCGCATCTCGCAAAACATCATTCATGATGGCCCTCTGCCTGTCTTCTATCTATGAAGATCTCAAGATGTATCAGGTGGAGTTCAAAACAATGAACGCCAAGCTGCTGATGGACCCCAAGAGACAGATCTTCCTGGACCAGAACATGCTGGCAGTGATCGATGAGCTGATGCAAGCCTTGAACTTCAACTCAGAGACAGTGCCGCAAAAGTCCTCGTTGGAGGAACCAGATTTTTACAAAACCAAAATCAAGCTGTGTATCCTTCTTCACGCCTTTCGGATCAGAGCCGTGACTATCGACCGGGTGATGTCATACCTGAATGCTTCC  13 hIL12AB_009ATGTGCCACCAGCAGCTGGTCATCAGCTGGTTTAGCCTGGTCTTCCTGGCCAGCCC ORFCCTGGTGGCCATCTGGGAGCTGAAGAAAGATGTCTATGTTGTAGAGCTGGACTGGTACCCAGATGCTCCTGGAGAAATGGTGGTTCTCACCTGCGACACGCCAGAAGAAGATGGCATCACCTGGACGCTGGACCAGAGCAGCGAAGTACTGGGCAGTGGAAAAACGCTGACCATACAAGTAAAAGAATTTGGCGATGCTGGCCAGTACACCTGCCACAAAGGAGGAGAAGTACTGAGCCACAGCCTGCTGCTGCTGCACAAGAAAGAAGATGGCATCTGGAGCACCGACATTTTAAAAGACCAGAAGGAGCCCAAGAACAAAACCTTCCTTCGATGTGAGGCGAAGAACTACAGTGGCCGCTTCACCTGCTGGTGGCTCACCACCATCAGCACCGACCTCACCTTCTCGGTGAAGAGCAGCCGTGGTAGCTCAGACCCCCAAGGAGTCACCTGTGGGGCGGCCACGCTGTCGGCAGAAAGAGTTCGAGGCGACAACAAGGAATATGAATACTCGGTGGAATGTCAAGAAGACTCGGCCTGCCCGGCGGCAGAAGAAAGTCTGCCCATAGAAGTCATGGTGGATGCTGTTCACAAATTAAAATATGAAAACTACACCAGCAGCTTCTTCATCAGAGACATCATCAAGCCAGACCCCCCCAAGAACCTGCAGCTGAAGCCCCTGAAGAACAGCAGACAAGTGGAAGTTTCCTGGGAGTACCCAGACACGTGGAGCACGCCGCACAGCTACTTCAGCCTCACCTTCTGTGTACAAGTACAAGGCAAGAGCAAGAGAGAGAAGAAAGATCGTGTCTTCACCGACAAAACCTCGGCGACGGTCATCTGCAGGAAGAATGCAAGCATCTCGGTTCGAGCCCAGGACCGCTACTACAGCAGCAGCTGGAGTGAGTGGGCCTCGGTGCCCTGCAGTGGTGGCGGCGGCGGCGGCAGCAGAAACCTTCCTGTGGCCACGCCGGACCCTGGCATGTTTCCGTGCCTGCACCACAGCCAAAATTTATTACGAGCTGTTAGCAACATGCTGCAGAAAGCAAGACAAACTTTAGAATTCTACCCCTGCACCTCAGAAGAAATAGACCATGAAGACATCACCAAAGATAAAACCAGCACTGTAGAGGCCTGCCTGCCCCTGGAGCTCACCAAGAACGAGAGCTGCCTCAATAGCAGAGAGACCAGCTTCATCACCAATGGCAGCTGCCTGGCCAGCAGGAAAACCAGCTTCATGATGGCGCTCTGCCTGAGCAGCATCTATGAAGATCTGAAGATGTACCAAGTAGAATTTAAAACCATGAATGCCAAGCTGCTCATGGACCCCAAGAGACAAATATTCCTCGACCAAAACATGCTGGCTGTCATTGATGAGCTCATGCAAGCATTAAACTTCAACTCAGAGACGGTGCCCCAGAAGAGCAGCCTGGAGGAGCCAGACTTCTACAAAACCAAGATCAAGCTCTGCATCTTATTACATGCCTTCCGCATCCGGGCGGTCACCATTGACCGTGTCATGTCCTACTTAAATGCCAGC  14 hIL12AB_010ATGTGCCACCAGCAGCTTGTCATCTCCTGGTTTTCTCTTGTCTTCCTCGCTTCTCC ORFTCTTGTGGCCATCTGGGAGCTGAAGAAAGATGTCTATGTTGTAGAGCTGGACTGGTACCCGGACGCTCCTGGAGAAATGGTGGTTCTCACCTGCGACACTCCTGAAGAAGATGGCATCACCTGGACGCTGGACCAAAGCAGCGAAGTTTTAGGCTCTGGAAAAACGCTGACCATACAAGTAAAAGAATTTGGCGACGCTGGCCAGTACACGTGCCACAAAGGAGGAGAAGTTTTAAGCCACAGTTTACTTCTTCTTCACAAGAAAGAAGATGGCATCTGGAGTACGGACATTTTAAAAGACCAGAAGGAGCCTAAGAACAAAACCTTCCTCCGCTGTGAAGCTAAGAACTACAGTGGTCGTTTCACCTGCTGGTGGCTCACCACCATCTCCACTGACCTCACCTTCTCTGTAAAATCAAGCCGTGGTTCTTCTGACCCCCAAGGAGTCACCTGTGGGGCTGCCACGCTCAGCGCTGAAAGAGTTCGAGGCGACAACAAGGAATATGAATATTCTGTGGAATGTCAAGAAGATTCTGCCTGCCCGGCGGCAGAAGAAAGTCTTCCCATAGAAGTCATGGTGGACGCTGTTCACAAATTAAAATATGAAAACTACACCAGCAGCTTCTTCATTCGTGACATCATCAAACCAGACCCTCCTAAGAACCTTCAGTTAAAACCGCTGAAGAACAGCAGACAAGTGGAAGTTTCCTGGGAGTACCCGGACACGTGGAGTACGCCGCACTCCTACTTCAGTTTAACCTTCTGTGTACAAGTACAAGGAAAATCAAAAAGAGAGAAGAAAGATCGTGTCTTCACTGACAAAACATCTGCCACGGTCATCTGCCGTAAGAACGCTTCCATCTCGGTTCGAGCCCAGGACCGCTACTACAGCAGCAGCTGGAGTGAGTGGGCATCTGTTCCCTGCAGTGGTGGCGGCGGCGGCGGCAGCCGCAACCTTCCTGTGGCCACGCCGGACCCTGGCATGTTCCCGTGCCTTCACCACTCGCAAAATCTTCTTCGTGCTGTTTCTAACATGCTGCAGAAGGCGAGACAAACTTTAGAATTCTACCCGTGCACTTCTGAAGAAATAGACCATGAAGACATCACCAAGGACAAAACCAGCACGGTGGAGGCCTGCCTTCCTTTAGAACTTACTAAGAACGAAAGTTGCCTTAACAGCCGTGAGACCAGCTTCATCACCAATGGCAGCTGCCTTGCTAGCAGGAAGACCAGCTTCATGATGGCGCTGTGCCTTTCTTCCATCTATGAAGATCTTAAGATGTACCAAGTAGAATTTAAAACCATGAATGCCAAATTATTAATGGACCCCAAGAGACAAATATTCCTCGACCAAAACATGCTGGCTGTCATTGATGAGCTCATGCAAGCATTAAACTTCAACTCAGAAACTGTTCCCCAGAAGTCATCTTTAGAAGAACCGGACTTCTACAAAACAAAAATAAAACTCTGCATTCTTCTTCATGCCTTCCGCATCCGTGCTGTCACCATTGACCGTGTCATGTCCTACTTAAATGCTTCT  15 hIL12AB_011ATGTGCCACCAGCAGCTGGTGATCAGCTGGTTCAGCCTGGTGTTCCTGGCCAGCCC ORFCCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGCTGGACTGGTACCCGGACGCGCCGGGGGAGATGGTGGTGCTGACGTGCGACACGCCGGAGGAGGACGGGATCACGTGGACGCTGGACCAGAGCAGCGAGGTGCTGGGGAGCGGGAAGACGCTGACGATCCAGGTGAAGGAGTTCGGGGACGCGGGGCAGTACACGTGCCACAAGGGGGGGGAGGTGCTGAGCCACAGCCTGCTGCTGCTGCACAAGAAGGAGGACGGGATCTGGAGCACGGACATCCTGAAGGACCAGAAGGAGCCGAAGAACAAGACGTTCCTGAGGTGCGAGGCGAAGAACTACAGCGGGAGGTTCACGTGCTGGTGGCTGACGACGATCAGCACGGACCTGACGTTCAGCGTGAAGAGCAGCAGGGGGAGCAGCGACCCGCAGGGGGTGACGTGCGGGGCGGCGACGCTGAGCGCGGAGAGGGTGAGGGGGGACAACAAGGAGTACGAGTACAGCGTGGAGTGCCAGGAGGACAGCGCGTGCCCGGCGGCGGAGGAGAGCCTGCCGATCGAGGTGATGGTGGACGCGGTGCACAAGCTGAAGTACGAGAACTACACGAGCAGCTTCTTCATCAGGGACATCATCAAGCCGGACCCGCCGAAGAACCTGCAGCTGAAGCCGCTGAAGAACAGCAGGCAGGTGGAGGTGAGCTGGGAGTACCCGGACACGTGGAGCACGCCGCACAGCTACTTCAGCCTGACGTTCTGCGTGCAGGTGCAGGGGAAGAGCAAGAGGGAGAAGAAGGACAGGGTGTTCACGGACAAGACGAGCGCGACGGTGATCTGCAGGAAGAACGCGAGCATCAGCGTGAGGGCGCAGGACAGGTACTACAGCAGCAGCTGGAGCGAGTGGGCGAGCGTGCCGTGCAGCGGGGGGGGGGGGGGGGGGAGCAGGAACCTGCCGGTGGCGACGCCGGACCCGGGGATGTTCCCGTGCCTGCACCACAGCCAGAACCTGCTGAGGGCGGTGAGCAACATGCTGCAGAAGGCGAGGCAGACGCTGGAGTTCTACCCGTGCACGAGCGAGGAGATCGACCACGAGGACATCACGAAGGACAAGACGAGCACGGTGGAGGCGTGCCTGCCGCTGGAGCTGACGAAGAACGAGAGCTGCCTGAACAGCAGGGAGACGAGCTTCATCACGAACGGGAGCTGCCTGGCGAGCAGGAAGACGAGCTTCATGATGGCGCTGTGCCTGAGCAGCATCTACGAGGACCTGAAGATGTACCAGGTGGAGTTCAAGACGATGAACGCGAAGCTGCTGATGGACCCGAAGAGGCAGATCTTCCTGGACCAGAACATGCTGGCGGTGATCGACGAGCTGATGCAGGCGCTGAACTTCAACAGCGAGACGGTGCCGCAGAAGAGCAGCCTGGAGGAGCCGGACTTCTACAAGACGAAGATCAAGCTGTGCATCCTGCTGCACGCGTTCAGGATCAGGGCGGTGACGATCGACAGGGTGATGAGCTACCTGAACGCGAGC  16 hIL12AB_012ATGTGCCATCAGCAGCTGGTGATCAGCTGGTTCAGCCTCGTGTTTCTGGCCAGCCC ORFCCTGGTGGCCATTTGGGAACTCAAGAAGGACGTGTATGTAGTGGAACTCGACTGGTACCCTGACGCCCCAGGCGAAATGGTGGTCTTAACCTGCGACACCCCTGAGGAGGACGGAATCACCTGGACCTTGGACCAGAGCTCCGAGGTCCTCGGCAGTGGCAAGACCCTGACCATACAGGTGAAAGAATTTGGAGACGCAGGGCAATACACATGTCACAAGGGCGGGGAGGTTCTTTCTCACTCCCTTCTGCTTCTACATAAAAAGGAAGACGGAATTTGGTCTACCGACATCCTCAAGGACCAAAAGGAGCCTAAGAATAAAACCTTCTTACGCTGTGAAGCTAAAAACTACAGCGGCAGATTCACTTGCTGGTGGCTCACCACCATTTCTACCGACCTGACCTTCTCGGTGAAGTCTTCAAGGGGCTCTAGTGATCCACAGGGAGTGACATGCGGGGCCGCCACACTGAGCGCTGAACGGGTGAGGGGCGATAACAAGGAGTATGAATACTCTGTCGAGTGTCAGGAGGATTCAGCTTGTCCCGCAGCTGAAGAGTCACTCCCCATAGAGGTTATGGTCGATGCTGTGCATAAACTGAAGTACGAAAACTACACCAGCAGCTTCTTCATTCGGGACATTATAAAACCTGACCCCCCCAAGAACCTGCAACTTAAACCCCTGAAAAACTCTCGGCAGGTCGAAGTTAGCTGGGAGTACCCTGATACTTGGTCCACCCCCCACTCGTACTTCTCACTGACTTTCTGTGTGCAGGTGCAGGGCAAGAGCAAGAGAGAGAAAAAAGATCGTGTATTCACAGACAAGACCTCTGCCACCGTGATCTGCAGAAAAAACGCTTCCATCAGTGTCAGAGCCCAAGACCGGTACTATAGTAGTAGCTGGAGCGAGTGGGCAAGTGTCCCCTGCTCTGGCGGCGGAGGGGGCGGCTCTCGAAACCTCCCCGTCGCTACCCCTGATCCAGGAATGTTCCCTTGCCTGCATCACTCACAGAATCTGCTGAGAGCGGTCAGCAACATGCTGCAGAAAGCTAGGCAAACACTGGAGTTTTATCCTTGTACCTCAGAGGAGATCGACCACGAGGATATTACCAAGGACAAGACCAGCACGGTGGAGGCCTGCTTGCCCCTGGAACTGACAAAGAATGAATCCTGCCTTAATAGCCGTGAGACCTCTTTTATAACAAACGGATCCTGCCTGGCCAGCAGGAAGACCTCCTTCATGATGGCCCTCTGCCTGTCCTCAATCTACGAAGACCTGAAGATGTACCAGGTGGAATTTAAAACTATGAACGCCAAGCTGTTGATGGACCCCAAGCGGCAGATCTTTCTGGATCAAAATATGCTGGCTGTGATCGACGAACTGATGCAGGCCCTCAACTTTAACAGCGAGACCGTGCCACAAAAGAGCAGTCTTGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATCCTCCTTCATGCCTTCAGGATAAGAGCTGTCACCATCGACAGAGTCATGAGTTACCTGAATGCATCC  17 hIL12AB_013ATGTGCCACCAGCAGCTGGTCATCTCCTGGTTCAGTCTTGTCTTCCTGGCCTCGCC ORFGCTGGTGGCCATCTGGGAGCTGAAGAAAGATGTTTATGTTGTAGAGCTGGACTGGTACCCAGATGCTCCTGGAGAAATGGTGGTCCTCACCTGTGACACGCCAGAAGAAGATGGCATCACCTGGACGCTGGACCAGAGCAGTGAAGTTCTTGGAAGTGGAAAAACGCTGACCATACAAGTAAAAGAATTTGGAGATGCTGGCCAGTACACCTGCCACAAAGGAGGAGAAGTTCTCAGCCACAGTTTATTATTACTTCACAAGAAAGAAGATGGCATCTGGTCCACGGACATTTTAAAAGACCAGAAGGAGCCCAAAAATAAAACATTTCTTCGATGTGAGGCCAAGAACTACAGTGGTCGTTTCACCTGCTGGTGGCTGACCACCATCTCCACAGACCTCACCTTCAGTGTAAAAAGCAGCCGTGGTTCTTCTGACCCCCAAGGAGTCACCTGTGGGGCTGCCACGCTCTCTGCAGAAAGAGTTCGAGGGGACAACAAAGAATATGAGTACTCGGTGGAATGTCAAGAAGACTCGGCCTGCCCAGCTGCTGAGGAGAGTCTTCCCATAGAAGTCATGGTGGATGCTGTTCACAAATTAAAATATGAAAACTACACCAGCAGCTTCTTCATCAGAGACATCATCAAACCTGACCCGCCCAAGAACTTACAGCTGAAGCCGCTGAAAAACAGCAGACAAGTAGAAGTTTCCTGGGAGTACCCGGACACCTGGTCCACGCCGCACTCCTACTTCTCCCTCACCTTCTGTGTACAAGTACAAGGCAAGAGCAAGAGAGAGAAGAAAGATCGTGTCTTCACGGACAAAACATCAGCCACGGTCATCTGCAGGAAAAATGCCAGCATCTCGGTGCGGGCCCAGGACCGCTACTACAGCAGCAGCTGGAGTGAGTGGGCATCTGTGCCCTGCAGTGGTGGTGGGGGTGGTGGCAGCAGAAACCTTCCTGTGGCCACTCCAGACCCTGGCATGTTCCCGTGCCTTCACCACTCCCAAAATTTACTTCGAGCTGTTTCTAACATGCTGCAGAAAGCAAGACAAACTTTAGAATTCTACCCGTGCACTTCTGAAGAAATTGACCATGAAGACATCACAAAAGATAAAACCAGCACAGTGGAGGCCTGTCTTCCTTTAGAGCTGACCAAAAATGAATCCTGCCTCAACAGCAGAGAGACCAGCTTCATCACCAATGGCAGCTGCCTGGCCTCCAGGAAAACCAGCTTCATGATGGCGCTCTGCCTCAGCTCCATCTATGAAGATTTGAAGATGTACCAAGTAGAATTTAAAACCATGAATGCCAAATTATTAATGGACCCCAAGAGGCAGATATTTTTAGATCAAAACATGCTGGCAGTTATTGATGAGCTCATGCAAGCATTAAACTTCAACAGTGAGACTGTACCTCAAAAAAGCAGCCTTGAAGAGCCGGACTTCTACAAAACCAAGATCAAACTCTGCATTTTACTTCATGCCTTCCGCATCCGGGCGGTCACCATTGACCGTGTCATGTCCTACTTAAATGCCTCG  18 hIL12AB_014ATGTGCCACCAGCAGCTTGTGATTTCTTGGTTCTCTCTTGTGTTCCTTGCTTCTCC ORFTCTTGTGGCTATTTGGGAGTTAAAAAAGGACGTGTACGTGGTGGAGCTTGACTGGTACCCTGATGCTCCTGGCGAGATGGTGGTGCTTACTTGTGACACTCCTGAGGAGGACGGCATTACTTGGACTCTTGACCAGTCTTCTGAGGTGCTTGGCTCTGGCAAGACTCTTACTATTCAGGTGAAGGAGTTCGGGGATGCTGGCCAGTACACTTGCCACAAGGGCGGCGAGGTGCTTTCTCACTCTCTTCTTCTTCTTCACAAGAAGGAGGACGGCATTTGGTCTACTGACATTTTAAAAGACCAGAAGGAGCCCAAGAACAAGACTTTCCTTCGTTGCGAGGCCAAGAACTACTCTGGCCGTTTCACTTGCTGGTGGCTTACTACTATTTCTACTGACCTTACTTTCTCTGTGAAGTCTTCTCGTGGCTCTTCTGACCCTCAGGGCGTGACTTGTGGGGCTGCTACTCTTTCTGCTGAGCGTGTGCGTGGGGACAACAAGGAGTACGAGTACTCTGTGGAGTGCCAGGAGGACTCTGCTTGCCCTGCTGCTGAGGAGTCTCTTCCTATTGAGGTGATGGTGGATGCTGTGCACAAGTTAAAATACGAGAACTACACTTCTTCTTTCTTCATTCGTGACATTATTAAGCCTGACCCTCCCAAGAACCTTCAGTTAAAACCTTTAAAAAACTCTCGTCAGGTGGAGGTGTCTTGGGAGTACCCTGACACTTGGTCTACTCCTCACTCTTACTTCTCTCTTACTTTCTGCGTGCAGGTGCAGGGCAAGTCTAAGCGTGAGAAGAAGGACCGTGTGTTCACTGACAAGACTTCTGCTACTGTGATTTGCAGGAAGAATGCATCTATTTCTGTGCGTGCTCAGGACCGTTACTACTCTTCTTCTTGGTCTGAGTGGGCTTCTGTGCCTTGCTCTGGCGGCGGCGGCGGCGGCTCTAGAAATCTTCCTGTGGCTACTCCTGACCCTGGCATGTTCCCTTGCCTTCACCACTCTCAGAACCTTCTTCGTGCTGTGAGCAACATGCTTCAGAAGGCTCGTCAGACTCTTGAGTTCTACCCTTGCACTTCTGAGGAGATTGACCACGAGGACATCACCAAGGACAAGACTTCTACTGTGGAGGCTTGCCTTCCTCTTGAGCTTACCAAGAATGAATCTTGCTTAAATTCTCGTGAGACTTCTTTCATCACCAACGGCTCTTGCCTTGCCTCGCGCAAGACTTCTTTCATGATGGCTCTTTGCCTTTCTTCTATTTACGAGGACTTAAAAATGTACCAGGTGGAGTTCAAGACTATGAATGCAAAGCTTCTTATGGACCCCAAGCGTCAGATTTTCCTTGACCAGAACATGCTTGCTGTGATTGACGAGCTTATGCAGGCTTTAAATTTCAACTCTGAGACTGTGCCTCAGAAGTCTTCTCTTGAGGAGCCTGACTTCTACAAGACCAAGATTAAGCTTTGCATTCTTCTTCATGCTTTCCGTATTCGTGCTGTGACTATTGACCGTGTGATGTCTTACTTAAATGCTTCT  19 hIL12AB_015ATGTGTCACCAGCAGCTGGTGATCAGCTGGTTTAGCCTGGTGTTTCTGGCCAGCCC ORFCCTGGTGGCCATATGGGAACTGAAGAAAGATGTGTATGTGGTAGAACTGGATTGGTATCCGGATGCCCCCGGCGAAATGGTGGTGCTGACCTGTGACACCCCCGAAGAAGATGGTATCACCTGGACCCTGGACCAGAGCAGCGAGGTGCTGGGCAGCGGCAAAACCCTGACCATCCAAGTGAAAGAGTTTGGCGATGCCGGCCAGTACACCTGTCACAAAGGCGGCGAGGTGCTAAGCCATTCGCTGCTGCTGCTGCACAAAAAGGAAGATGGCATCTGGAGCACCGATATCCTGAAGGACCAGAAAGAACCCAAAAATAAGACCTTTCTAAGATGCGAGGCCAAGAATTATAGCGGCCGTTTCACCTGCTGGTGGCTGACGACCATCAGCACCGATCTGACCTTCAGCGTGAAAAGCAGCAGAGGCAGCAGCGACCCCCAAGGCGTGACGTGCGGCGCCGCCACCCTGAGCGCCGAGAGAGTGAGAGGCGACAACAAGGAGTATGAGTACAGCGTGGAGTGCCAGGAGGACAGCGCCTGCCCCGCCGCCGAGGAGAGCCTGCCCATCGAGGTGATGGTGGATGCCGTGCACAAGCTGAAGTATGAAAACTACACCAGCAGCTTCTTCATCAGAGACATCATCAAACCCGACCCCCCCAAGAACCTGCAGCTGAAGCCCCTGAAGAATAGCAGACAGGTGGAGGTGAGCTGGGAGTACCCCGACACCTGGAGCACCCCCCATAGCTACTTCAGCCTGACCTTCTGCGTGCAGGTGCAGGGCAAGAGCAAGAGAGAAAAGAAAGATAGAGTGTTCACGGACAAGACCAGCGCCACGGTGATCTGCAGAAAAAATGCCAGCATCAGCGTGAGAGCCCAGGACAGATACTATAGCAGCAGCTGGAGCGAATGGGCCAGCGTGCCCTGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCTGCCCGTGGCCACCCCCGACCCCGGCATGTTCCCCTGCCTGCACCACAGCCAAAACCTGCTGAGAGCCGTGAGCAACATGCTGCAGAAGGCCAGACAAACCCTGGAATTTTACCCCTGCACCAGCGAAGAGATCGATCATGAAGATATCACCAAAGATAAAACCAGCACCGTGGAGGCCTGTCTGCCCCTGGAACTGACCAAGAATGAGAGCTGCCTAAATAGCAGAGAGACCAGCTTCATAACCAATGGCAGCTGCCTGGCCAGCAGAAAGACCAGCTTTATGATGGCCCTGTGCCTGAGCAGCATCTATGAAGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAATGCCAAGCTGCTGATGGATCCCAAGAGACAGATCTTTCTGGATCAAAACATGCTGGCCGTGATCGATGAGCTGATGCAGGCCCTGAATTTCAACAGCGAGACCGTGCCCCAAAAAAGCAGCCTGGAAGAACCGGATTTTTATAAAACCAAAATCAAGCTGTGCATACTGCTGCATGCCTTCAGAATCAGAGCCGTGACCATCGATAGAGTGATGAGCTATCTGAATGCCAGC  20 hIL12AB_016ATGTGCCACCAGCAGCTGGTCATCAGCTGGTTCAGCCTGGTCTTCCTGGCCAGCCC ORFCCTGGTGGCCATCTGGGAGCTGAAGAAGGATGTTTATGTTGTGGAGCTGGACTGGTACCCAGATGCCCCTGGGGAGATGGTGGTGCTGACCTGTGACACCCCAGAAGAGGATGGCATCACCTGGACCCTGGACCAGAGCTCAGAAGTGCTGGGCAGTGGAAAAACCCTGACCATCCAGGTGAAGGAGTTTGGAGATGCTGGCCAGTACACCTGCCACAAGGGTGGTGAAGTGCTGAGCCACAGCCTGCTGCTGCTGCACAAGAAGGAGGATGGCATCTGGAGCACAGACATCCTGAAGGACCAGAAGGAGCCCAAGAACAAGACCTTCCTTCGCTGTGAAGCCAAGAACTACAGTGGCCGCTTCACCTGCTGGTGGCTGACCACCATCAGCACAGACCTCACCTTCTCGGTGAAGAGCAGCAGAGGCAGCTCAGACCCCCAGGGTGTCACCTGTGGGGCGGCCACGCTGTCGGCGGAGAGAGTTCGAGGGGACAACAAGGAGTATGAATACTCGGTGGAGTGCCAGGAGGACTCGGCGTGCCCGGCGGCAGAAGAGAGCCTGCCCATAGAAGTGATGGTGGATGCTGTGCACAAGCTGAAGTATGAAAACTACACCAGCAGCTTCTTCATCAGAGACATCATCAAGCCAGACCCGCCCAAGAACCTGCAGCTGAAGCCCCTGAAGAACAGCAGACAAGTGGAGGTTTCCTGGGAGTACCCAGACACGTGGAGCACCCCCCACAGCTACTTCAGCCTGACCTTCTGTGTCCAGGTGCAGGGCAAGAGCAAGAGAGAGAAGAAGGACAGAGTCTTCACAGACAAGACCTCGGCCACGGTCATCTGCAGAAAGAATGCCTCCATCTCGGTTCGAGCCCAGGACAGATACTACAGCAGCAGCTGGTCAGAATGGGCCTCGGTGCCCTGCAGTGGTGGCGGCGGCGGCGGCAGCAGAAACCTGCCTGTTGCCACCCCAGACCCTGGGATGTTCCCCTGCCTGCACCACAGCCAGAACTTATTACGAGCTGTTTCTAACATGCTGCAGAAGGCCAGACAAACCCTGGAGTTCTACCCCTGCACCTCAGAAGAGATTGACCATGAAGACATCACCAAGGACAAGACCAGCACTGTAGAGGCCTGCCTGCCCCTGGAGCTGACCAAGAATGAAAGCTGCCTGAACAGCAGAGAGACCAGCTTCATCACCAATGGAAGCTGCCTGGCCAGCAGAAAGACCAGCTTCATGATGGCCCTGTGCCTGAGCAGCATCTATGAAGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAATGCAAAGCTGCTGATGGACCCCAAGAGACAAATATTTTTGGACCAGAACATGCTGGCTGTCATTGATGAGCTGATGCAGGCCCTGAACTTCAACTCAGAAACTGTACCCCAGAAGAGCAGCCTGGAGGAGCCAGACTTCTACAAGACCAAGATCAAGCTGTGCATCCTGCTTCATGCTTTCAGAATCAGAGCTGTCACCATTGACCGCGTGATGAGCTACTTAAATGCCTCG  21 hIL12AB_017ATGTGCCACCAGCAGCTGGTAATCAGCTGGTTTTCCCTCGTCTTTCTGGCATCACC ORFCCTGGTGGCTATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGCTGGATTGGTACCCTGACGCCCCGGGGGAAATGGTGGTGTTAACATGCGACACGCCTGAGGAGGACGGCATCACCTGGACACTGGACCAGAGCAGCGAGGTGCTTGGGTCTGGTAAAACTCTGACTATTCAGGTGAAAGAGTTCGGGGATGCCGGCCAATATACTTGCCACAAGGGTGGCGAGGTGCTTTCTCATTCTCTGCTCCTGCTGCACAAGAAAGAAGATGGCATTTGGTCTACTGATATTCTGAAAGACCAGAAGGAGCCCAAGAACAAGACCTTTCTGAGATGCGAGGCTAAAAACTACAGCGGAAGATTTACCTGCTGGTGGCTGACCACAATCTCAACCGACCTGACATTTTCAGTGAAGTCCAGCAGAGGGAGCTCCGACCCTCAGGGCGTGACCTGCGGAGCCGCCACTCTGTCCGCAGAAAGAGTGAGAGGTGATAATAAGGAGTACGAGTATTCAGTCGAGTGCCAAGAGGACTCTGCCTGCCCAGCCGCCGAGGAGAGCCTGCCAATCGAGGTGATGGTAGATGCGGTACACAAGCTGAAGTATGAGAACTACACATCCTCCTTCTTCATAAGAGACATTATCAAGCCTGACCCACCTAAAAATCTGCAACTCAAGCCTTTGAAAAATTCAAGACAGGTGGAGGTGAGCTGGGAGTACCCTGATACTTGGAGCACCCCCCATAGCTACTTTTCGCTGACATTCTGCGTCCAGGTGCAGGGCAAGTCAAAGAGAGAGAAGAAGGATCGCGTGTTCACTGATAAGACAAGCGCCACAGTGATCTGCAGAAAAAACGCTAGCATTAGCGTCAGAGCACAGGACCGGTATTACTCCAGCTCCTGGAGCGAATGGGCATCTGTGCCCTGCAGCGGTGGGGGCGGAGGCGGATCTAGAAACCTCCCCGTTGCCACACCTGATCCTGGAATGTTCCCCTGTCTGCACCACAGCCAGAACCTGCTGAGAGCAGTGTCTAACATGCTCCAGAAGGCCAGGCAGACCCTGGAGTTTTACCCCTGCACCAGCGAGGAAATCGATCACGAGGACATCACCAAAGATAAAACCTCCACCGTGGAGGCCTGCCTGCCCCTGGAACTGACCAAAAACGAGAGCTGCCTGAATAGCAGGGAGACCTCCTTCATCACCAACGGCTCATGCCTTGCCAGCCGGAAAACTAGCTTCATGATGGCCCTGTGCCTGTCTTCGATCTATGAGGACCTGAAAATGTACCAGGTCGAATTTAAGACGATGAACGCAAAGCTGCTGATGGACCCCAAGCGGCAGATCTTTCTGGACCAGAACATGCTGGCAGTCATAGATGAGTTGATGCAGGCATTAAACTTGAACAGCGAGACCGTGCCTCAGAAGTCCAGCCTCGAGGAGCCAGATTTTTATAAGACCAAGATCAAACTATGCATCCTGCTGCATGCTTTCAGGATTAGAGCCGTCACCATCGATCGAGTCATGTCTTACCTGAATGCTAGC  22 hIL12AB_018ATGTGTCACCAACAGTTAGTAATCTCCTGGTTTTCTCTGGTGTTTCTGGCCAGCCC ORFCCTCGTGGCCATCTGGGAGCTTAAAAAGGATGTGTACGTGGTGGAGCTGGACTGGTATCCCGATGCACCAGGCGAAATGGTCGTGCTGACCTGCGATACCCCTGAAGAAGATGGCATCACCTGGACTCTGGACCAGTCTTCCGAGGTGCTTGGATCTGGCAAGACTCTGACAATACAAGTTAAGGAGTTCGGGGACGCAGGACAGTACACCTGCCACAAAGGCGGCGAGGTCCTGAGTCACTCCCTGTTACTGCTCCACAAGAAAGAGGACGGCATTTGGTCCACCGACATTCTGAAGGACCAGAAGGAGCCTAAGAATAAAACTTTCCTGAGATGCGAGGCAAAAAACTATAGCGGCCGCTTTACTTGCTGGTGGCTTACAACAATCTCTACCGATTTAACTTTCTCCGTGAAGTCTAGCAGAGGATCCTCTGACCCGCAAGGAGTGACTTGCGGAGCCGCCACCTTGAGCGCCGAAAGAGTCCGTGGCGATAACAAAGAATACGAGTACTCCGTGGAGTGCCAGGAAGATTCCGCCTGCCCAGCTGCCGAGGAGTCCCTGCCCATTGAAGTGATGGTGGATGCCGTCCACAAGCTGAAGTACGAAAACTATACCAGCAGCTTCTTCATCCGGGATATCATTAAGCCCGACCCTCCTAAAAACCTGCAACTTAAGCCCCTAAAGAATAGTCGGCAGGTTGAGGTCAGCTGGGAATATCCTGACACATGGAGCACCCCCCACTCTTATTTCTCCCTGACCTTCTGCGTGCAGGTGCAGGGCAAGAGTAAACGGGAGAAAAAGGACAGGGTCTTTACCGATAAAACCAGCGCTACGGTTATCTGTCGGAAGAACGCTTCCATCTCCGTCCGCGCTCAGGATCGTTACTACTCGTCCTCATGGAGCGAGTGGGCCAGCGTGCCCTGCAGCGGCGGCGGTGGAGGCGGATCCAGAAATCTGCCTGTTGCCACACCAGACCCTGGCATGTTCCCCTGTCTGCATCATAGCCAGAACCTGCTCAGAGCCGTGAGCAACATGCTCCAGAAGGCCAGGCAGACATTGGAGTTCTACCCGTGTACATCTGAGGAAATCGATCACGAAGATATAACCAAGGACAAAACCTCTACAGTAGAGGCTTGTTTGCCCCTGGAGTTGACCAAAAACGAGAGTTGCCTGAACAGTCGCGAGACAAGCTTCATTACTAACGGCAGCTGTCTCGCCTCCAGAAAGACATCCTTCATGATGGCCCTGTGTCTTTCCAGCATATACGAAGACCTGAAAATGTACCAGGTCGAGTTCAAAACAATGAACGCCAAGCTGCTTATGGACCCCAAGAGACAGATCTTCCTCGACCAAAACATGCTCGCTGTGATCGATGAGCTGATGCAGGCTCTCAACTTCAATTCCGAAACAGTGCCACAGAAGTCCAGTCTGGAAGAACCCGACTTCTACAAGACCAAGATTAAGCTGTGTATTTTGCTGCATGCGTTTAGAATCAGAGCCGTGACCATTGATCGGGTGATGAGCTACCTGAACGCCTCG  23 hIL12AB_019ATGTGCCACCAGCAGCTTGTCATCTCCTGGTTTTCTCTTGTCTTCCTGGCCTCGCC ORFGCTGGTGGCCATCTGGGAGCTGAAGAAAGATGTCTATGTTGTAGAGCTGGACTGGTACCCAGATGCTCCTGGAGAAATGGTGGTTCTCACCTGTGACACTCCTGAAGAAGATGGCATCACCTGGACGCTGGACCAAAGCTCAGAAGTTCTTGGCAGTGGAAAAACGCTGACCATACAAGTAAAAGAATTTGGGGATGCTGGCCAGTACACGTGCCACAAAGGAGGAGAAGTTCTCAGCCACAGTTTACTTCTTCTTCACAAGAAAGAAGATGGCATCTGGTCCACGGACATTTTAAAAGACCAGAAGGAGCCCAAGAACAAAACCTTCCTCCGCTGTGAGGCCAAGAACTACAGTGGTCGTTTCACCTGCTGGTGGCTCACCACCATCTCCACTGACCTCACCTTCTCTGTAAAAAGCAGCCGTGGTTCTTCTGACCCCCAAGGAGTCACCTGTGGGGCTGCCACGCTCTCGGCAGAAAGAGTTCGAGGGGACAACAAGGAATATGAATATTCTGTGGAATGTCAAGAAGATTCTGCCTGCCCGGCGGCAGAAGAAAGTCTTCCCATAGAAGTCATGGTGGATGCTGTTCACAAATTAAAATATGAAAACTACACCAGCAGCTTCTTCATTCGTGACATCATCAAACCAGACCCGCCCAAGAACCTTCAGTTAAAACCTTTAAAAAACAGCAGACAAGTAGAAGTTTCCTGGGAGTACCCGGACACGTGGTCCACGCCGCACTCCTACTTCAGTTTAACCTTCTGTGTACAAGTACAAGGAAAATCAAAAAGAGAGAAGAAAGATCGTGTCTTCACTGACAAAACATCTGCCACGGTCATCTGCAGGAAGAATGCCTCCATCTCGGTTCGAGCCCAGGACCGCTACTACAGCAGCAGCTGGAGTGAGTGGGCATCTGTTCCCTGCAGTGGTGGCGGCGGCGGCGGCAGCCGCAACCTTCCTGTGGCCACGCCGGACCCTGGCATGTTCCCGTGCCTTCACCACTCCCAAAATCTTCTTCGTGCTGTTTCTAACATGCTGCAGAAGGCGCGCCAAACTTTAGAATTCTACCCGTGCACTTCTGAAGAAATAGACCATGAAGACATCACCAAAGATAAAACCAGCACGGTGGAGGCCTGCCTTCCTTTAGAGCTGACCAAGAATGAATCCTGCCTCAACAGCAGAGAGACCAGCTTCATCACCAATGGCAGCTGCCTGGCCTCGCGCAAGACCAGCTTCATGATGGCGCTGTGCCTTTCTTCCATCTATGAAGATTTAAAGATGTACCAAGTAGAATTTAAAACCATGAATGCCAAATTATTAATGGACCCCAAAAGACAAATATTTTTGGATCAAAACATGCTGGCTGTCATTGATGAGCTCATGCAAGCATTAAACTTCAACTCAGAAACTGTTCCCCAGAAGTCATCTTTAGAAGAGCCGGACTTCTACAAAACAAAAATAAAACTCTGCATTCTTCTTCATGCCTTCCGCATCCGTGCTGTCACCATTGACCGTGTCATGTCCTACTTAAATGCTTCT  24 hIL12AB_020ATGTGCCACCAGCAGCTGGTGATCAGCTGGTTCAGCCTGGTGTTCCTGGCTAGCCC ORFTCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGTTAGACTGGTACCCCGACGCTCCCGGCGAGATGGTGGTGCTGACCTGCGACACCCCCGAGGAGGACGGGATCACCTGGACCCTGGATCAGTCAAGCGAGGTGCTGGGAAGCGGCAAGACCCTGACCATCCAGGTGAAGGAGTTCGGCGACGCCGGCCAATACACTTGCCACAAGGGAGGCGAGGTGCTGTCCCACTCCCTCCTGCTGCTGCACAAAAAGGAAGACGGCATCTGGAGCACCGACATCCTGAAAGACCAGAAGGAGCCTAAGAACAAGACATTCCTCAGATGCGAGGCCAAGAATTACTCCGGGAGATTCACCTGTTGGTGGCTGACCACCATCAGCACAGACCTGACCTTCAGCGTGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGTGACCTGTGGCGCCGCCACCCTGAGCGCCGAAAGAGTGCGCGGCGACAACAAGGAGTACGAGTACTCCGTGGAATGCCAGGAGGACAGCGCCTGCCCCGCCGCCGAGGAGAGCCTGCCCATCGAGGTGATGGTGGACGCCGTCCACAAGCTGAAGTACGAGAACTACACCTCTAGCTTCTTCATCCGGGACATCATCAAGCCCGATCCCCCCAAGAACCTGCAGCTGAAACCCCTGAAGAACAGCAGACAGGTGGAGGTGAGCTGGGAGTATCCCGACACCTGGTCCACCCCCCACAGCTATTTTAGCCTGACCTTCTGCGTGCAAGTGCAGGGCAAGAGCAAGAGAGAGAAGAAGGACCGCGTGTTCACCGACAAAACCAGCGCCACCGTGATCTGCAGAAAGAACGCCAGCATCAGCGTGAGGGCCCAGGATAGATACTACAGTTCCAGCTGGAGCGAGTGGGCCAGCGTGCCCTGCAGCGGCGGCGGCGGGGGAGGCTCTAGAAACCTGCCCGTGGCTACCCCCGATCCCGGAATGTTCCCCTGCCTGCACCACAGCCAGAACCTGCTGAGGGCGGTGTCCAACATGCTTCAGAAGGCCCGGCAGACCCTGGAGTTCTACCCCTGTACCTCTGAGGAGATCGATCATGAGGACATCACAAAGGACAAAACCAGCACCGTGGAGGCCTGCCTGCCCCTGGAGCTGACCAAGAACGAGAGCTGCCTGAACTCCCGCGAGACCAGCTTCATCACGAACGGCAGCTGCCTGGCCAGCAGGAAGACCTCCTTCATGATGGCCCTGTGCCTGAGCAGCATCTACGAGGACCTGAAAATGTACCAGGTGGAGTTTAAGACCATGAACGCCAAGCTGCTGATGGACCCCAAGCGGCAAATCTTCCTGGACCAGAACATGCTGGCAGTGATCGACGAGCTCATGCAGGCCCTGAACTTCAATAGCGAGACAGTCCCCCAGAAGAGCAGCCTGGAGGAGCCCGACTTTTACAAGACCAAGATCAAGCTGTGCATCCTGCTGCACGCCTTTAGAATCCGTGCCGTGACCATTGACAGAGTGATGAGCTACCTGAATGCCAGC  25 hIL12AB_021ATGTGCCACCAGCAGCTGGTGATCAGCTGGTTCAGCCTGGTGTTCCTGGCCAGCCC ORFTCTGGTTGCCATCTGGGAGCTGAAGAAAGACGTGTACGTCGTGGAACTGGACTGGTATCCGGACGCCCCGGGCGAGATGGTGGTGCTGACCTGTGACACCCCCGAGGAGGACGGCATCACCTGGACGCTGGACCAATCCTCCGAGGTGCTGGGAAGCGGCAAGACCCTGACCATCCAGGTGAAGGAATTCGGGGACGCCGGGCAGTACACCTGCCACAAGGGGGGCGAAGTGCTGTCCCACTCGCTGCTGCTCCTGCATAAGAAGGAGGATGGAATCTGGTCCACCGACATCCTCAAAGATCAGAAGGAGCCCAAGAACAAGACGTTCCTGCGCTGTGAAGCCAAGAATTATTCGGGGCGATTCACGTGCTGGTGGCTGACAACCATCAGCACCGACCTGACGTTTAGCGTGAAGAGCAGCAGGGGGTCCAGCGACCCCCAGGGCGTGACGTGCGGCGCCGCCACCCTCTCCGCCGAGAGGGTGCGGGGGGACAATAAGGAGTACGAGTACAGCGTGGAATGCCAGGAGGACAGCGCCTGCCCCGCCGCGGAGGAAAGCCTCCCGATAGAGGTGATGGTGGACGCCGTGCACAAGCTCAAGTATGAGAATTACACCAGCAGCTTTTTCATCCGGGACATTATCAAGCCCGACCCCCCGAAGAACCTCCAGCTGAAGCCCCTGAAGAACAGCCGGCAGGTGGAAGTCTCCTGGGAGTATCCCGACACCTGGAGCACCCCGCACAGCTACTTCTCCCTGACCTTCTGTGTGCAGGTGCAGGGCAAGTCCAAGAGGGAAAAGAAGGACAGGGTTTTCACCGACAAGACCAGCGCGACCGTGATCTGCCGGAAGAACGCCAGCATAAGCGTCCGCGCCCAAGATAGGTACTACAGCAGCTCCTGGAGCGAGTGGGCTAGCGTGCCCTGCAGCGGGGGCGGGGGTGGGGGCTCCAGGAACCTGCCAGTGGCGACCCCCGACCCCGGCATGTTCCCCTGCCTCCATCACAGCCAGAACCTGCTGAGGGCCGTCAGCAATATGCTGCAGAAGGCCAGGCAGACCCTGGAATTCTACCCCTGCACGTCGGAGGAGATCGATCACGAGGATATCACAAAAGACAAGACTTCCACCGTGGAGGCCTGCCTGCCCCTGGAGCTCACCAAGAATGAGTCCTGTCTGAACTCCCGGGAAACCAGCTTCATCACCAACGGGTCCTGCCTGGCCAGCAGGAAGACCAGCTTTATGATGGCCCTGTGCCTGTCGAGCATCTACGAGGACCTGAAGATGTACCAGGTCGAGTTCAAGACAATGAACGCCAAGCTGCTGATGGACCCCAAGAGGCAAATCTTCCTGGACCAGAATATGCTTGCCGTCATCGACGAGCTCATGCAGGCCCTGAACTTCAACTCCGAGACCGTGCCCCAGAAGAGCAGCCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATCCTGCTGCACGCGTTCAGGATCCGGGCAGTCACCATCGACCGTGTGATGTCCTACCTGAACGCCAGC  26 hIL12AB_022ATGTGCCATCAGCAGCTGGTGATCAGCTGGTTCAGCCTGGTGTTCCTCGCCTCTCC ORFCCTGGTGGCCATCTGGGAGCTCAAAAAGGACGTGTACGTGGTGGAGCTCGACTGGTACCCAGACGCCCCCGGGGAGATGGTGGTGCTGACCTGCGACACCCCCGAAGAAGACGGCATCACGTGGACCCTCGACCAGTCCAGCGAGGTGCTGGGGAGCGGGAAGACTCTGACCATCCAGGTCAAGGAGTTCGGGGACGCCGGGCAGTACACGTGCCACAAGGGCGGCGAAGTCTTAAGCCACAGCCTGCTCCTGCTGCACAAGAAGGAGGACGGGATCTGGTCCACAGACATACTGAAGGACCAGAAGGAGCCGAAGAATAAAACCTTTCTGAGGTGCGAGGCCAAGAACTATTCCGGCAGGTTCACGTGCTGGTGGCTTACAACAATCAGCACAGACCTGACGTTCAGCGTGAAGTCCAGCCGCGGCAGCAGCGACCCCCAGGGGGTGACCTGCGGCGCCGCCACCCTGAGCGCCGAGCGGGTGCGCGGGGACAACAAGGAGTACGAGTACTCCGTGGAGTGCCAGGAAGACAGCGCCTGTCCCGCCGCCGAAGAGAGCCTGCCTATCGAGGTCATGGTAGATGCAGTGCATAAGCTGAAGTACGAGAACTATACGAGCAGCTTTTTCATACGCGACATCATCAAGCCCGACCCCCCCAAGAACCTGCAGCTTAAGCCCCTGAAGAATAGCCGGCAGGTGGAGGTCTCCTGGGAGTACCCCGACACCTGGTCAACGCCCCACAGCTACTTCTCCCTGACCTTTTGTGTCCAAGTCCAGGGAAAGAGCAAGAGGGAGAAGAAAGATCGGGTGTTCACCGACAAGACCTCCGCCACGGTGATCTGCAGGAAGAACGCCAGCATCTCCGTGAGGGCGCAAGACAGGTACTACTCCAGCAGCTGGTCCGAATGGGCCAGCGTGCCCTGCTCCGGCGGCGGGGGCGGCGGCAGCCGAAACCTACCCGTGGCCACGCCGGATCCCGGCATGTTTCCCTGCCTGCACCACAGCCAGAACCTCCTGAGGGCCGTGTCCAACATGCTGCAGAAGGCCAGGCAGACTCTGGAGTTCTACCCCTGCACGAGCGAGGAGATCGATCACGAGGACATCACCAAGGATAAGACCAGCACTGTGGAGGCCTGCCTTCCCCTGGAGCTGACCAAGAACGAGAGCTGTCTGAACTCCAGGGAGACCTCATTCATCACCAACGGCTCCTGCCTGGCCAGCAGGAAAACCAGCTTCATGATGGCCTTGTGTCTCAGCTCCATCTACGAGGACCTGAAGATGTATCAGGTCGAGTTCAAGACAATGAACGCCAAGCTGCTGATGGACCCCAAAAGGCAGATCTTCCTGGACCAGAACATGCTGGCCGTCATCGACGAGCTGATGCAGGCCCTGAACTTCAACAGCGAGACGGTGCCCCAGAAAAGCTCCCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATCCTGCTGCACGCCTTCAGGATCAGGGCAGTGACCATCGACCGGGTGATGTCATACCTTAACGCCAGC  27 hIL12AB_023ATGTGCCATCAGCAGCTGGTGATCTCCTGGTTCAGCCTGGTGTTTCTGGCCTCGCC ORFCCTGGTCGCCATCTGGGAGCTGAAGAAAGACGTGTACGTCGTCGAACTGGACTGGTACCCCGACGCCCCCGGGGAGATGGTGGTGCTGACCTGCGACACGCCGGAGGAGGACGGCATCACCTGGACCCTGGATCAAAGCAGCGAGGTGCTGGGCAGCGGCAAGACCCTGACCATCCAAGTGAAGGAATTCGGCGATGCCGGCCAGTACACCTGTCACAAAGGGGGCGAGGTGCTCAGCCACAGCCTGCTGCTGCTGCACAAGAAGGAGGATGGCATCTGGAGCACCGATATCCTGAAGGACCAGAAAGAGCCCAAGAACAAGACGTTCCTGAGGTGCGAGGCCAAGAACTACAGCGGTAGGTTCACGTGTTGGTGGCTGACCACCATCAGCACCGACCTGACGTTCAGCGTGAAGAGCTCCAGGGGCAGCTCCGACCCACAGGGGGTGACGTGCGGGGCCGCAACCCTCAGCGCCGAAAGGGTGCGGGGGGACAACAAGGAGTACGAATACTCCGTGGAGTGCCAGGAAGATTCGGCCTGCCCCGCCGCGGAGGAGAGCCTCCCCATCGAGGTAATGGTGGACGCCGTGCATAAGCTGAAGTACGAGAACTACACCAGCTCGTTCTTCATCCGAGACATCATCAAACCCGACCCGCCCAAAAATCTGCAGCTCAAGCCCCTGAAGAACTCCAGGCAGGTGGAGGTGAGCTGGGAGTACCCCGACACCTGGTCCACCCCGCACAGCTACTTCTCCCTGACATTCTGCGTGCAGGTGCAGGGCAAGAGCAAGCGGGAGAAGAAGGACAGGGTGTTCACCGACAAGACGAGCGCCACCGTGATCTGCCGAAAGAACGCCAGCATCTCGGTGCGCGCCCAGGATAGGTACTATTCCAGCTCCTGGAGCGAGTGGGCCTCGGTACCCTGCAGCGGCGGCGGGGGCGGCGGCAGTAGGAATCTGCCCGTGGCTACCCCGGACCCGGGCATGTTCCCCTGCCTCCACCACAGCCAGAACCTGCTGAGGGCCGTGAGCAACATGCTGCAGAAGGCCAGACAGACGCTGGAGTTCTACCCCTGCACGAGCGAGGAGATCGACCACGAGGACATCACCAAGGATAAAACTTCCACCGTCGAGGCCTGCCTGCCCTTGGAGCTGACCAAGAATGAATCCTGTCTGAACAGCAGGGAGACCTCGTTTATCACCAATGGCAGCTGCCTCGCCTCCAGGAAGACCAGCTTCATGATGGCCCTCTGTCTGAGCTCCATCTATGAGGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAACGCGAAGCTGCTGATGGACCCCAAGAGGCAGATCTTCCTGGATCAGAATATGCTGGCGGTGATCGACGAGCTCATGCAGGCCCTCAATTTCAATAGCGAGACAGTGCCCCAGAAGTCCTCCCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGTATCCTGCTGCACGCCTTCCGGATCCGGGCCGTCACCATCGACCGGGTCATGAGCTACCTCAATGCCAGC  28 hIL12AB_024ATGTGCCACCAGCAGCTGGTGATCTCCTGGTTCTCCCTGGTGTTCCTGGCCTCGCC ORFCCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTCGTGGAGCTCGACTGGTACCCCGACGCCCCTGGCGAGATGGTGGTGCTGACCTGCGACACCCCAGAGGAGGATGGCATCACCTGGACCCTGGATCAGTCCTCCGAGGTGCTGGGCTCCGGCAAGACGCTGACCATCCAAGTGAAGGAGTTCGGTGACGCCGGACAGTATACCTGCCATAAGGGCGGCGAGGTCCTGTCCCACAGCCTCCTCCTCCTGCATAAGAAGGAGGACGGCATCTGGAGCACCGACATCCTGAAGGACCAGAAGGAGCCCAAGAACAAGACCTTTCTGAGGTGCGAGGCCAAGAACTACAGCGGCCGATTCACCTGCTGGTGGCTCACCACCATATCCACCGACCTGACTTTCTCCGTCAAGTCCTCCCGGGGGTCCAGCGACCCCCAGGGAGTGACCTGCGGCGCCGCCACCCTCAGCGCCGAGCGGGTGCGGGGGGACAACAAGGAGTACGAATACTCCGTCGAGTGCCAGGAGGACTCCGCCTGCCCGGCCGCCGAGGAGAGCCTGCCCATCGAGGTGATGGTCGACGCGGTGCACAAGCTGAAGTACGAGAACTACACCAGCAGTTTCTTCATCAGGGATATCATCAAGCCAGATCCCCCGAAGAATCTGCAACTGAAGCCGCTGAAAAACTCACGACAGGTGGAGGTGAGCTGGGAGTACCCCGACACGTGGAGCACCCCACATTCCTACTTCAGCCTGACCTTCTGCGTGCAGGTCCAGGGCAAGAGCAAGCGGGAGAAGAAGGACAGGGTGTTCACGGATAAGACCAGTGCCACCGTGATCTGCAGGAAGAACGCCTCTATTAGCGTGAGGGCCCAGGATCGGTATTACTCCTCGAGCTGGAGCGAATGGGCCTCCGTGCCCTGCAGTGGGGGGGGTGGAGGCGGGAGCAGGAACCTGCCCGTAGCAACCCCCGACCCCGGGATGTTCCCCTGTCTGCACCACTCGCAGAACCTGCTGCGCGCGGTGAGCAACATGCTCCAAAAAGCCCGTCAGACCTTAGAGTTCTACCCCTGCACCAGCGAAGAAATCGACCACGAAGACATCACCAAGGACAAAACCAGCACCGTGGAGGCGTGCCTGCCGCTGGAGCTGACCAAGAACGAGAGCTGCCTCAACTCCAGGGAGACCAGCTTTATCACCAACGGCTCGTGCCTAGCCAGCCGGAAAACCAGCTTCATGATGGCCCTGTGCCTGAGCTCCATTTACGAGGACCTGAAGATGTATCAGGTGGAGTTCAAGACCATGAATGCCAAACTCCTGATGGACCCCAAGAGGCAGATCTTCCTGGACCAGAACATGCTCGCGGTGATCGATGAGCTGATGCAGGCCCTGAACTTTAATAGCGAGACCGTGCCCCAGAAAAGCAGCCTGGAGGAGCCGGACTTCTACAAGACCAAAATCAAGCTGTGCATCCTGCTCCACGCCTTCCGCATCCGGGCCGTGACCATCGACAGGGTGATGAGCTACCTGAACGCCAGC  29 hIL12AB_025ATGTGCCATCAGCAGCTGGTGATTTCCTGGTTCTCCCTGGTGTTCCTGGCCAGCCC ORFCCTCGTGGCGATCTGGGAGCTAAAGAAGGACGTGTACGTGGTGGAGCTGGACTGGTACCCGGACGCACCCGGCGAGATGGTCGTTCTGACCTGCGATACGCCAGAGGAGGACGGCATCACCTGGACCCTCGATCAGAGCAGCGAGGTCCTGGGGAGCGGAAAGACCCTGACCATCCAGGTCAAGGAGTTCGGCGACGCCGGCCAGTACACCTGCCACAAAGGTGGCGAGGTCCTGAGCCACTCGCTGCTGCTCCTGCATAAGAAGGAGGACGGAATCTGGAGCACAGACATCCTGAAAGACCAGAAGGAGCCCAAGAACAAGACCTTCCTGAGGTGCGAGGCCAAGAACTACAGCGGGCGCTTCACGTGCTGGTGGCTGACCACCATCAGCACGGACCTCACCTTCTCCGTGAAGAGCAGCCGGGGATCCAGCGATCCCCAAGGCGTCACCTGCGGCGCGGCCACCCTGAGCGCGGAGAGGGTCAGGGGCGATAATAAGGAGTATGAGTACAGCGTGGAGTGCCAGGAGGACAGCGCCTGCCCGGCCGCCGAGGAGTCCCTGCCAATCGAAGTGATGGTCGACGCCGTGCACAAGCTGAAGTACGAGAACTACACCAGCAGCTTCTTCATCCGGGATATCATCAAGCCCGATCCCCCGAAGAACCTGCAGCTGAAGCCCCTCAAGAACAGCCGGCAGGTGGAGGTGAGTTGGGAGTACCCCGACACCTGGTCAACGCCCCACAGCTACTTCTCCCTGACCTTCTGTGTGCAGGTGCAGGGAAAGAGCAAGAGGGAGAAGAAAGACCGGGTCTTCACCGACAAGACCAGCGCCACGGTGATCTGCAGGAAGAACGCAAGCATCTCCGTGAGGGCCCAGGACAGGTACTACAGCTCCAGCTGGTCCGAATGGGCCAGCGTGCCCTGTAGCGGCGGCGGGGGCGGTGGCAGCCGCAACCTCCCAGTGGCCACCCCCGACCCCGGCATGTTCCCCTGCCTGCACCACAGCCAGAATCTGCTGAGGGCCGTGAGTAACATGCTGCAGAAGGCAAGGCAAACCCTCGAATTCTATCCCTGCACCTCCGAGGAGATCGACCACGAGGATATCACCAAGGACAAGACCAGCACCGTCGAGGCCTGTCTCCCCCTGGAGCTGACCAAGAATGAGAGCTGCCTGAACAGCCGGGAGACCAGCTTCATCACCAACGGGAGCTGCCTGGCCTCCAGGAAGACCTCGTTCATGATGGCGCTGTGCCTCTCAAGCATATACGAGGATCTGAAGATGTACCAGGTGGAGTTTAAGACGATGAACGCCAAGCTGCTGATGGACCCGAAGAGGCAGATCTTCCTGGACCAGAACATGCTGGCCGTGATAGACGAGCTCATGCAGGCCCTGAACTTCAACTCCGAGACCGTGCCGCAGAAGTCATCCCTCGAGGAGCCCGACTTCTATAAGACCAAGATCAAGCTGTGCATCCTGCTCCACGCCTTCCGGATAAGGGCCGTGACGATCGACAGGGTGATGAGCTACCTTAACGCCAGC  30 hIL12AB_026ATGTGCCACCAGCAGCTCGTGATCAGCTGGTTCTCCCTGGTGTTTCTCGCCAGCCC ORFCCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGCTGGACTGGTACCCTGACGCCCCGGGGGAGATGGTCGTGCTGACCTGCGACACCCCCGAAGAGGACGGTATCACCTGGACCCTGGACCAGTCCAGCGAGGTGCTGGGCAGCGGCAAGACCCTGACTATTCAAGTCAAGGAGTTCGGAGACGCCGGCCAGTACACCTGCCACAAGGGTGGAGAGGTGTTATCACACAGCCTGCTGCTGCTGCACAAGAAGGAAGACGGGATCTGGAGCACCGACATCCTGAAGGACCAGAAGGAGCCCAAAAACAAGACCTTCCTGCGGTGCGAGGCCAAGAACTATTCGGGCCGCTTTACGTGCTGGTGGCTGACCACCATCAGCACTGATCTCACCTTCAGCGTGAAGTCCTCCCGGGGGTCGTCCGACCCCCAGGGGGTGACCTGCGGGGCCGCCACCCTGTCCGCCGAGAGAGTGAGGGGCGATAATAAGGAGTACGAGTACAGCGTTGAGTGCCAGGAAGATAGCGCCTGTCCCGCCGCCGAGGAGAGCCTGCCCATCGAGGTGATGGTGGACGCCGTCCACAAGCTGAAGTATGAGAACTACACCTCAAGCTTCTTCATCAGGGACATCATCAAACCCGATCCGCCCAAGAATCTGCAGCTGAAGCCCCTGAAAAATAGCAGGCAGGTGGAGGTGAGCTGGGAGTACCCCGACACCTGGTCCACCCCCCATAGCTATTTCTCCCTGACGTTCTGCGTGCAGGTGCAAGGGAAGAGCAAGCGGGAGAAGAAGGACCGGGTGTTCACCGACAAGACCTCCGCCACCGTGATCTGTAGGAAGAACGCGTCGATCTCGGTCAGGGCCCAGGACAGGTATTACAGCAGCAGCTGGAGCGAGTGGGCGAGCGTGCCCTGCTCGGGCGGCGGCGGCGGCGGGAGCAGAAATCTGCCCGTGGCCACCCCAGACCCCGGAATGTTCCCCTGCCTGCACCATTCGCAGAACCTCCTGAGGGCCGTGAGCAACATGCTGCAGAAGGCCCGCCAGACGCTGGAGTTCTACCCCTGCACGAGCGAGGAGATCGACCACGAAGACATCACCAAGGACAAAACCAGCACCGTGGAGGCCTGCCTGCCCCTGGAGCTGACCAAAAACGAATCCTGCCTCAACAGCCGGGAGACCAGCTTCATCACCAACGGCAGCTGCCTGGCCAGCCGAAAGACCTCCTTCATGATGGCCCTCTGCCTGAGCAGCATCTATGAGGATCTGAAGATGTATCAGGTGGAGTTCAAGACCATGAATGCCAAGCTGCTGATGGACCCCAAGAGGCAGATATTCCTGGACCAGAATATGCTGGCCGTGATCGACGAGCTGATGCAGGCCCTGAACTTCAACAGCGAGACCGTCCCCCAGAAGTCCAGCCTGGAGGAGCCGGACTTTTACAAAACGAAGATCAAGCTGTGCATACTGCTGCACGCCTTCAGGATCCGGGCCGTGACAATCGACAGGGTGATGTCCTACCTGAACGCCAGC  31 hIL12AB_027ATGTGTCACCAGCAGCTGGTGATCAGCTGGTTCTCCCTGGTGTTCCTGGCCAGCCC ORFCCTGGTGGCCATCTGGGAGCTCAAGAAGGACGTCTACGTCGTGGAGCTGGATTGGTACCCCGACGCTCCCGGGGAGATGGTGGTGCTGACCTGCGACACCCCCGAGGAGGACGGCATCACCTGGACGCTGGACCAGAGCTCAGAGGTGCTGGGAAGCGGAAAGACACTGACCATCCAGGTGAAGGAGTTCGGGGATGCCGGGCAGTATACCTGCCACAAGGGCGGCGAAGTGCTGAGCCATTCCCTGCTGCTGCTGCACAAGAAGGAGGACGGCATATGGTCCACCGACATCCTGAAGGATCAGAAGGAGCCGAAGAATAAAACCTTCCTGAGGTGCGAGGCCAAGAATTACAGCGGCCGATTCACCTGCTGGTGGCTGACCACCATCAGCACCGACCTGACCTTCAGTGTGAAGTCCTCACGGGGCAGCTCAGATCCCCAGGGCGTGACCTGCGGGGCCGCGACACTCAGCGCCGAGCGGGTGAGGGGTGATAACAAGGAGTACGAGTATTCTGTGGAGTGCCAGGAAGACTCCGCCTGTCCCGCCGCCGAGGAGTCCCTGCCCATCGAGGTGATGGTGGACGCCGTGCATAAACTGAAGTACGAGAACTACACCTCCAGCTTCTTCATCCGGGATATAATCAAGCCCGACCCTCCGAAAAACCTGCAGCTGAAGCCCCTTAAAAACAGCCGGCAGGTGGAGGTGAGCTGGGAGTACCCCGACACCTGGAGCACCCCCCATAGCTATTTCAGCCTGACCTTCTGCGTGCAGGTGCAGGGGAAGTCCAAGCGCGAGAAAAAGGACCGGGTGTTCACCGACAAGACGAGCGCCACCGTGATCTGCCGGAAGAACGCCAGTATAAGCGTAAGGGCCCAGGATAGGTACTACAGCTCCAGCTGGTCGGAGTGGGCCTCCGTGCCCTGTTCCGGCGGCGGGGGGGGTGGCAGCAGGAACCTCCCCGTGGCCACGCCGGACCCCGGCATGTTCCCGTGCCTGCACCACTCCCAAAACCTCCTGCGGGCCGTCAGCAACATGCTGCAAAAGGCGCGGCAGACCCTGGAGTTTTACCCCTGTACCTCCGAAGAGATCGAGGAGGAGGATATGAGCAAGGATAAGACCTCCACCGTGGAGGCCTGTCTCCCCCTGGAGCTGACCAAGAACGAGAGCTGTCTTAACAGCAGAGAGACCTCGTTCATAACGAACGGCTCCTGCCTCGCTTCCAGGAAGACGTCGTTCATGATGGCGCTGTGCCTGTCCAGCATCTACGAGGACCTGAAGATGTATCAGGTCGAGTTCAAAACCATGAACGCCAAGCTGCTGATGGACCCCAAGAGGCAGATCTTCCTGGACCAGAACATGCTCGCCGTGATCGACGAGCTGATGCAGGCCCTGAACTTCAACAGCGAAACCGTGCCCCAGAAGTCAAGCCTGGAGGAGCCGGACTTCTATAAGACCAAGATCAAGCTGTGTATCCTGCTACACGCTTTTCGTATCCGGGCCGTGACCATCGACAGGGTTATGTCGTACTTGAACGCCAGC  32 hIL12AB_028ATGTGCCACCAACAGCTCGTGATCAGCTGGTTCAGCCTGGTGTTCCTGGCCAGCCC ORFGCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGCTGGACTGGTACCCCGACGCCCCCGGCGAGATGGTGGTCCTGACCTGCGACACGCCGGAAGAGGACGGCATCACCTGGACCCTGGATCAGTCCAGCGAGGTGCTGGGCTCCGGCAAGACCCTGACCATTCAGGTGAAGGAGTTCGGCGACGCCGGTCAGTACACCTGCCACAAGGGCGGCGAGGTGCTGAGCCACAGCCTACTGCTCCTGCACAAAAAGGAGGATGGAATCTGGTCCACCGACATCCTCAAGGACCAGAAGGAGCCGAAGAACAAGACGTTCCTCCGGTGCGAGGCCAAGAACTACAGCGGCAGGTTTACCTGCTGGTGGCTGACCACCATCAGCACCGACCTGACATTTTCCGTGAAGAGCAGCCGCGGCAGCAGCGATCCCCAGGGCGTGACCTGCGGGGCGGCCACCCTGTCCGCCGAGCGTGTGAGGGGCGACAACAAGGAGTACGAGTACAGCGTGGAATGCCAGGAGGACAGCGCCTGTCCCGCCGCCGAGGAGAGCCTGCCAATCGAGGTCATGGTGGACGCCGTGCACAAGCTGAAGTACGAGAACTACACGAGCAGCTTCTTCATCAGGGACATCATCAAACCGGACCCGCCCAAGAACCTGCAGCTGAAACCCTTGAAAAACAGCAGGCAGGTGGAAGTGTCTTGGGAGTACCCCGACACCTGGTCCACCCCCCACAGCTACTTTAGCCTGACCTTCTGTGTGCAGGTCCAGGGCAAGTCCAAGAGGGAGAAGAAGGACAGGGTGTTCACCGACAAAACCAGCGCCACCGTGATCTGCAGGAAGAACGCCTCCATCAGCGTGCGGGCCCAGGACAGGTATTACAGCTCGTCGTGGAGCGAGTGGGCCAGCGTGCCCTGCTCCGGGGGAGGCGGCGGCGGAAGCCGGAATCTGCCCGTGGCCACCCCCGATCCCGGCATGTTCCCGTGTCTGCACCACAGCCAGAACCTGCTGCGGGCCGTGAGCAACATGCTGCAGAAGGCCCGCCAAACCCTGGAGTTCTACCCCTGTACAAGCGAGGAGATCGACCATGAGGACATTACCAAGGACAAGACCAGCACCGTGGAGGCCTGCCTGCCCCTCGAGCTCACAAAGAACGAATCCTGCCTGAATAGCCGCGAGACCAGCTTTATCACGAACGGGTCCTGCCTCGCCAGCCGGAAGACAAGCTTCATGATGGCCCTGTGCCTGAGCAGCATCTACGAGGACCTGAAAATGTACCAAGTGGAGTTCAAAACGATGAACGCCAAGCTGCTGATGGACCCCAAGCGCCAGATCTTCCTGGACCAGAACATGCTGGCCGTCATCGACGAGCTCATGCAGGCCCTGAACTTCAACAGCGAGACCGTGCCCCAGAAGAGCAGCCTGGAGGAGCCCGACTTCTACAAGACGAAGATCAAGCTCTGCATCCTGCTGCACGCTTTCCGCATCCGCGCGGTGACCATCGACCGGGTGATGAGCTACCTCAACGCCAGT  33 hIL12AB_029ATGTGCCACCAACAGCTGGTGATCAGCTGGTTCAGCCTGGTGTTTCTGGCCTCCCC ORFTCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGCTGGACTGGTACCCTGACGCCCCCGGCGAAATGGTGGTGCTGACGTGCGACACCCCCGAGGAGGATGGCATCACCTGGACCCTGGACCAAAGCAGCGAGGTCCTCGGAAGCGGCAAGACCCTCACTATCCAAGTGAAGGAGTTCGGGGATGCGGGCCAGTACACCTGCCACAAGGGCGGCGAGGTGCTGTCTCATAGCCTGCTGCTCCTGCATAAGAAGGAAGACGGCATCTGGAGCACCGACATACTGAAGGATCAGAAGGAGCCCAAGAACAAGACCTTCCTGAGGTGCGAGGCCAAGAACTACTCCGGGCGCTTCACCTGTTGGTGGCTGACCACCATCTCCACCGACCTGACCTTCAGCGTGAAGAGCAGCAGGGGGAGCAGCGACCCCCAGGGGGTGACCTGCGGAGCCGCGACCTTGTCGGCCGAGCGGGTGAGGGGCGACAATAAGGAGTACGAGTACTCGGTCGAATGCCAGGAGGACTCCGCCTGCCCCGCCGCCGAGGAGTCCCTCCCCATCGAAGTGATGGTGGACGCCGTCCACAAGCTGAAGTACGAGAACTACACCAGCAGCTTCTTCATACGGGATATCATCAAGCCCGACCCCCCGAAGAACCTGCAGCTGAAACCCTTGAAGAACTCCAGGCAGGTGGAGGTGAGCTGGGAGTACCCCGACACCTGGTCCACCCCGCACTCATACTTCAGCCTGACCTTCTGTGTACAGGTCCAGGGCAAGAGCAAGAGGGAAAAGAAGGATAGGGTGTTCACCGACAAGACCTCCGCCACGGTGATCTGTCGGAAAAACGCCAGCATCTCCGTGCGGGCCCAGGACAGGTACTATTCCAGCAGCTGGAGCGAGTGGGCCTCCGTCCCCTGCTCCGGCGGCGGTGGCGGGGGCAGCAGGAACCTCCCCGTGGCCACCCCCGATCCCGGGATGTTCCCATGCCTGCACCACAGCCAAAACCTGCTGAGGGCCGTCTCCAATATGCTGCAGAAGGCGAGGCAGACCCTGGAGTTCTACCCCTGTACCTCCGAGGAGATCGACCACGAGGATATCACCAAGGACAAGACCTCCACGGTCGAGGCGTGCCTGCCCCTGGAGCTCACGAAGAACGAGAGCTGCCTTAACTCCAGGGAAACCTCGTTTATCACGAACGGCAGCTGCCTGGCGTCACGGAAGACCTCCTTTATGATGGCCCTATGTCTGTCCTCGATCTACGAGGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAACGCCAAGCTGCTGATGGATCCCAAGAGGCAGATTTTCCTGGACCAGAACATGCTGGCCGTGATTGACGAGCTGATGCAGGCGCTGAACTTCAACAGCGAGACAGTGCCGCAGAAGAGCTCCCTGGAGGAGCCGGACTTTTACAAGACCAAGATAAAGCTGTGCATCCTGCTCCACGCCTTCAGAATACGGGCCGTCACCATCGATAGGGTGATGTCTTACCTGAACGCCTCC  34 hIL12AB_030ATGTGCCACCAGCAGCTGGTGATTAGCTGGTTTAGCCTGGTGTTCCTGGCAAGCCC ORFCCTGGTGGCCATCTGGGAACTGAAAAAGGACGTGTACGTGGTCGAGCTGGATTGGTACCCCGACGCCCCCGGCGAAATGGTGGTGCTGACGTGTGATACCCCCGAGGAGGACGGGATCACCTGGACCCTGGATCAGAGCAGCGAGGTGCTGGGGAGCGGGAAGACCCTGACGATCCAGGTCAAGGAGTTCGGCGACGCTGGGCAGTACACCTGTCACAAGGGCGGGGAGGTGCTGTCCCACTCCCTGCTGCTCCTGCATAAGAAAGAGGACGGCATCTGGTCCACCGACATCCTCAAGGACCAGAAGGAGCCCAAGAACAAGACCTTCCTGCGGTGTGAGGCGAAGAACTACAGCGGCCGTTTCACCTGCTGGTGGCTGACGACAATCAGCACCGACTTGACGTTCTCCGTGAAGTCCTCCAGAGGCAGCTCCGACCCCCAAGGGGTGACGTGCGGCGCGGCCACCCTGAGCGCCGAGCGGGTGCGGGGGGACAACAAGGAGTACGAGTACTCCGTGGAGTGCCAGGAGGACAGCGCCTGTCCCGCAGCCGAGGAGTCCCTGCCCATCGAAGTCATGGTGGACGCCGTCCACAAGCTGAAGTACGAGAACTACACCAGCAGCTTCTTCATCCGCGATATCATCAAGCCCGATCCCCCCAAAAACCTGCAACTGAAGCCGCTGAAGAATAGCAGGCAGGTGGAGGTGTCCTGGGAGTACCCGGACACCTGGAGCACGCCCCACAGCTATTTCAGCCTGACCTTTTGCGTGCAGGTCCAGGGGAAGAGCAAGCGGGAGAAGAAGGACCGCGTGTTTACGGACAAAACCAGCGCCACCGTGATCTGCAGGAAGAACGCCAGCATCAGCGTGAGGGCCCAGGACAGGTACTACAGCAGCTCCTGGAGCGAGTGGGCCTCCGTGCCCTGTTCCGGAGGCGGCGGGGGCGGTTCCCGGAACCTCCCGGTGGCCACCCCCGACCCGGGCATGTTCCCGTGCCTGCACCACTCACAGAATCTGCTGAGGGCCGTGAGCAATATGCTGCAGAAGGCAAGGCAGACCCTGGAGTTTTATCCCTGCACCAGCGAGGAGATCGACCACGAAGACATCACCAAGGACAAGACCAGCACAGTGGAGGCCTGCCTGCCCCTGGAACTGACCAAGAACGAGTCCTGTCTGAACTCCCGGGAAACCAGCTTCATAACCAACGGCTCCTGTCTCGCCAGCAGGAAGACCAGCTTCATGATGGCCCTGTGCCTCAGCTCCATCTACGAGGACCTCAAGATGTACCAGGTTGAGTTCAAGACCATGAACGCCAAGCTCCTGATGGACCCCAAGAGGCAGATCTTCCTGGACCAGAATATGCTGGCCGTGATCGATGAGTTAATGCAGGCGCTGAACTTCAACAGCGAGACGGTGCCCCAAAAGTCCTCGCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATCCTCCTGCACGCCTTCCGAATCCGGGCCGTAACCATCGACAGGGTGATGAGCTATCTCAACGCCTCC  35 hIL12AB_031ATGTGCCACCAGCAGCTCGTGATCAGCTGGTTCTCGCTTGTGTTCCTGGCCTCCCC ORFCCTCGTCGCCATCTGGGAGCTGAAGAAAGACGTGTACGTGGTGGAGCTGGACTGGTATCCCGACGCCCCGGGGGAGATGGTGGTGCTGACCTGCGACACCCCGGAAGAGGACGGCATCACCTGGACGCTCGACCAGTCGTCCGAAGTGCTGGGGTCGGGCAAGACCCTCACCATCCAGGTGAAGGAGTTCGGAGACGCCGGCCAGTACACCTGTCATAAGGGGGGGGAGGTGCTGAGCCACAGCCTCCTGCTCCTGCACAAAAAGGAGGACGGCATCTGGAGCACCGATATCCTCAAGGACCAGAAGGAGCCCAAGAACAAGACGTTCCTGAGGTGTGAGGCCAAGAACTACAGCGGGCGGTTCACGTGTTGGTGGCTCACCACCATCTCCACCGACCTCACCTTCTCCGTGAAGTCAAGCAGGGGCAGCTCCGACCCCCAAGGCGTCACCTGCGGCGCCGCCACCCTGAGCGCCGAGAGGGTCAGGGGGGATAACAAGGAATACGAGTACAGTGTGGAGTGCCAAGAGGATAGCGCCTGTCCCGCCGCCGAAGAGAGCCTGCCCATCGAAGTGATGGTGGACGCCGTGCACAAGCTGAAGTACGAGAACTACACCTCCAGCTTCTTCATCAGGGATATCATCAAGCCCGATCCCCCCAAGAACCTGCAGCTGAAGCCCCTGAAGAACAGCAGGCAGGTGGAGGTGAGCTGGGAGTATCCCGACACGTGGAGCACCCCGCACAGCTACTTCTCGCTGACCTTCTGCGTGCAGGTGCAAGGGAAGTCCAAGAGGGAGAAGAAGGATAGGGTGTTCACCGACAAAACGAGCGCCACCGTGATCTGCCGGAAGAATGCCAGCATCTCTGTGAGGGCCCAGGACAGGTACTATTCCAGCTCCTGGTCGGAGTGGGCCAGCGTGCCCTGTAGCGGCGGGGGCGGGGGCGGCAGCAGGAACCTCCCGGTTGCCACCCCCGACCCCGGCATGTTTCCGTGCCTGCACCACTCGCAAAACCTGCTGCGCGCGGTCTCCAACATGCTGCAAAAAGCGCGCCAGACGCTGGAGTTCTACCCCTGCACCAGCGAGGAGATCGATCATGAAGATATCACCAAAGACAAGACCTCGACCGTGGAGGCCTGCCTGCCCCTGGAGCTCACCAAGAACGAAAGCTGCCTGAACAGCAGGGAGACAAGCTTCATCACCAACGGCAGCTGCCTGGCCTCCCGGAAGACCAGCTTCATGATGGCCCTGTGCCTGTCCAGCATCTACGAGGATCTGAAGATGTACCAAGTGGAGTTTAAGACCATGAACGCCAAGCTGTTAATGGACCCCAAAAGGCAGATCTTCCTGGATCAGAACATGCTGGCCGTCATCGACGAGCTGATGCAAGCCCTGAACTTCAACAGCGAGACGGTGCCCCAGAAGAGCAGCCTCGAGGAGCCCGACTTCTATAAGACCAAGATAAAGCTGTGCATTCTGCTGCACGCCTTCAGAATCAGGGCCGTGACCATCGATAGGGTGATGAGCTACCTGAACGCCAGC  36 hIL12AB_032ATGTGTCACCAGCAGCTGGTGATTTCCTGGTTCAGTCTGGTGTTTCTTGCCAGCCC ORFCCTGGTGGCCATCTGGGAGCTGAAGAAAGACGTATACGTCGTGGAGCTGGACTGGTATCCCGACGCTCCCGGCGAGATGGTGGTCCTCACCTGCGACACCCCAGAGGAGGACGGCATCACCTGGACCCTGGACCAGAGCTCCGAGGTCCTGGGCAGCGGTAAGACCCTCACCATCCAGGTGAAGGAGTTTGGTGATGCCGGGCAGTATACCTGCCACAAGGGCGGCGAGGTGCTGTCCCACAGCCTCCTGTTACTGCATAAGAAGGAGGATGGCATCTGGAGCACCGACATCCTCAAGGACCAGAAAGAGCCCAAGAACAAGACCTTTCTGCGGTGCGAGGCGAAAAATTACTCCGGCCGGTTCACCTGCTGGTGGCTGACCACCATCAGCACGGACCTGACGTTCTCCGTGAAGTCGAGCAGGGGGAGCTCCGATCCCCAGGGCGTGACCTGCGGCGCGGCCACCCTGAGCGCCGAGCGCGTCCGCGGGGACAATAAGGAATACGAATATAGCGTGGAGTGCCAGGAGGACAGCGCCTGCCCCGCGGCCGAGGAGAGCCTCCCGATCGAGGTGATGGTGGATGCCGTCCACAAGCTCAAATACGAAAACTACACCAGCAGCTTCTTCATTAGGGACATCATCAAGCCCGACCCCCCCAAAAACCTGCAGCTGAAGCCCCTGAAGAACAGCCGCCAGGTCGAGGTGTCATGGGAGTACCCAGACACCTGGAGCACCCCCCACTCCTACTTCAGCCTGACCTTCTGCGTCCAGGTGCAGGGAAAGTCCAAACGGGAGAAGAAGGATAGGGTCTTTACCGATAAGACGTCGGCCACCGTCATCTGCAGGAAGAACGCCAGCATAAGCGTGCGGGCGCAGGATCGGTACTACAGCTCGAGCTGGTCCGAATGGGCCTCCGTGCCCTGTAGCGGAGGGGGTGGCGGGGGCAGCAGGAACCTGCCCGTGGCCACCCCGGACCCGGGCATGTTTCCCTGCCTGCATCACAGTCAGAACCTGCTGAGGGCCGTGAGCAACATGCTCCAGAAGGCCCGCCAGACCCTGGAGTTTTACCCCTGCACCAGCGAAGAGATCGATGAGGAAGACATCACCAAAGACAAGACCTCCACCGTGGAGGCCTGTCTGCCCCTGGAGCTGACCAAGAACGAGAGCTGTCTGAACAGCAGGGAGACCTCCTTCATCACCAACGGCTCCTGCCTGGCATCCCGGAAGACCAGCTTCATGATGGCCCTGTGTCTGAGCTCTATCTACGAGGACCTGAAGATGTACCAGGTCGAGTTCAAGACCATGAACGCCAAGCTGCTGATGGACCCCAAGCGACAGATATTCCTGGACCAGAACATGCTCGCCGTGATCGATGAACTGATGCAAGCCCTGAACTTCAATAGCGAGACCGTGCCCCAGAAAAGCAGCCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAACTGTGCATACTGCTGCACGCGTTCAGGATCCGGGCCGTCACCATCGACCGGGTGATGTCCTATCTGAATGCCAGC  37 hIL12AB_033ATGTGCCACCAGCAGCTCGTGATTAGCTGGTTTTCGCTGGTGTTCCTGGCCAGCCC ORFTCTCGTGGCCATCTGGGAGCTGAAAAAAGACGTGTACGTGGTGGAGCTGGACTGGTACCCGGACGCCCCCGGCGAGATGGTGGTGCTGACGTGCGACACCCCGGAAGAGGACGGCATCACCTGGACCCTGGACCAGTCATCCGAGGTCCTGGGCAGCGGCAAGACGCTCACCATCCAGGTGAAGGAGTTCGGCGACGCCGGCCAGTACACATGCCATAAGGGCGGGGAGGTGCTGAGCCACAGCCTGCTCCTCCTGCACAAGAAGGAGGATGGCATCTGGTCTACAGACATCCTGAAGGACCAGAAAGAGCCCAAGAACAAGACCTTCCTCCGGTGCGAGGCCAAGAACTACTCCGGGCGGTTTACTTGTTGGTGGCTGACCACCATCAGCACCGACCTCACCTTCAGCGTGAAGAGCTCCCGAGGGAGCTCCGACCCCCAGGGGGTCACCTGCGGCGCCGCCACCCTGAGCGCCGAGCGGGTGAGGGGCGACAACAAGGAGTATGAATACAGCGTGGAATGCCAAGAGGACAGCGCCTGTCCCGCGGCCGAGGAAAGCCTGCCCATCGAGGTGATGGTGGACGCCGTCCACAAACTCAAGTACGAGAACTACACCAGCAGTTTCTTCATTCGCGACATCATCAAGCCGGACCCCCCCAAAAACCTGCAGCTCAAACCCCTGAAGAACAGCAGGCAGGTGGAGGTCAGCTGGGAGTACCCGGACACCTGGAGCACCCCCCATAGCTACTTCAGCCTGACCTTCTGCGTGCAGGTGCAGGGCAAGAGCAAACGCGAGAAGAAGGACCGGGTGTTTACCGACAAGACCAGCGCCACGGTGATCTGCCGAAAGAATGCAAGCATCTCCGTGAGGGCGCAGGACCGCTACTACTCTAGCAGCTGGAGCGAGTGGGCCAGCGTGCCCTGCAGCGGTGGCGGCGGAGGCGGCAGCCGTAACCTCCCCGTGGCCACCCCCGACCCCGGCATGTTCCCGTGTCTGCACCACTCCCAGAACCTGCTGAGGGCCGTCAGCAATATGCTGCAGAAGGCCCGGCAGACGCTGGAGTTCTACCCCTGCACCTCCGAGGAGATCGACCATGAGGACATTACCAAGGACAAGACGAGCACTGTGGAGGCCTGCCTGCCCCTGGAGCTCACCAAAAACGAGAGCTGCCTGAATAGCAGGGAGACGTCCTTCATCACCAACGGCAGCTGTCTGGCCAGCAGGAAGACCAGCTTCATGATGGCCCTGTGCCTCTCCTCCATATATGAGGATCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAACGCCAAGCTGCTGATGGATCCCAAGAGGCAGATCTTCCTGGACCAGAATATGCTGGCCGTGATTGACGAGCTGATGCAGGCCCTGAACTTTAATAGCGAGACCGTCCCCCAGAAGAGCAGCCTGGAGGAGCCCGACTTCTATAAGACCAAGATCAAGCTGTGCATACTGCTGCACGCGTTTAGGATAAGGGCCGTCACCATCGACAGGGTGATGAGCTACCTGAATGCCAGC  38 hIL12AB_034ATGTGCCACCAACAGCTGGTGATCTCCTGGTTCAGCCTGGTGTTCCTCGCCAGCCC ORFCCTGGTGGCCATCTGGGAGCTGAAGAAAGACGTGTACGTGGTGGAGCTGGACTGGTATCCCGACGCCCCCGGCGAGATGGTCGTGCTGACCTGCGACACCCCGGAGGAGGACGGCATCACCTGGACCCTGGATCAGTCCTCCGAGGTGCTGGGCAGCGGGAAGACCCTGACCATCCAGGTGAAAGAGTTCGGAGATGCCGGCCAGTATACCTGTCACAAGGGGGGTGAGGTGCTGAGCCATAGCCTCTTGCTTCTGCACAAGAAGGAGGACGGCATCTGGTCCACCGACATCCTCAAGGACCAAAAGGAGCCGAAGAATAAAACGTTCCTGAGGTGCGAAGCCAAGAACTATTCCGGACGGTTCACCTGCTGGTGGCTGACCACCATCAGCACCGACCTCACCTTCTCCGTAAAGTCAAGCAGGGGCAGCTCCGACCCCCAGGGCGTGACCTGCGGAGCCGCCACCCTGAGCGCAGAGAGGGTGAGGGGCGACAACAAGGAGTACGAATACTCCGTCGAGTGCCAGGAGGACAGCGCCTGCCCCGCCGCCGAGGAAAGTCTGCCCATCGAGGTGATGGTGGACGCCGTGCACAAGCTCAAATACGAGAACTACACCAGCAGCTTCTTCATCCGGGATATCATCAAGCCCGACCCTCCAAAGAATCTGCAGCTGAAACCCCTTAAGAACAGCAGGCAGGTGGAGGTCAGCTGGGAGTACCCCGACACCTGGAGCACGCCCCACTCCTACTTTAGCCTGACCTTTTGCGTGCAGGTGCAGGGGAAAAGCAAGCGGGAGAAGAAGGACAGGGTGTTCACCGATAAGACCTCCGCTACCGTGATCTGCAGGAAGAACGCCTCAATCAGCGTGAGGGCCCAGGATCGGTACTACTCCAGCTCCTGGAGCGAGTGGGCCAGCGTGCCCTGCTCTGGCGGTGGCGGCGGGGGCAGCCGGAACCTGCCGGTGGCCACTCCCGACCCGGGCATGTTCCCGTGCCTCCACCATTCCCAGAACCTGCTGCGGGCCGTGTCCAATATGCTCCAGAAGGCAAGGCAGACCCTGGAGTTCTACCCCTGCACCAGCGAGGAGATCGATCACGAGGACATCACCAAAGACAAAACCAGCACGGTCGAGGCCTGCCTGCCCCTGGAACTCACCAAGAACGAAAGCTGTCTCAACAGCCGCGAGACCAGCTTCATAACCAACGGTTCCTGTCTGGCCTCCCGCAAGACCAGCTTTATGATGGCCCTCTGTCTGAGCTCCATCTATGAAGACCTGAAAATGTACCAGGTGGAGTTCAAAACCATGAACGCCAAGCTTCTGATGGACCCCAAGAGGCAGATCTTCCTGGATCAGAACATGCTGGCCGTGATCGACGAGCTGATGCAGGCCCTGAACTTTAACTCCGAGACCGTGCCCCAGAAAAGCAGCCTGGAAGAGCCCGATTTCTACAAAACGAAGATCAAGCTGTGCATCCTGCTGCACGCCTTCCGGATCCGTGCGGTGACCATCGATAGGGTGATGAGCTACCTGAACGCCAGC  39 hIL12AB_035ATGTGCCACCAACAGCTGGTAATCAGCTGGTTCAGCCTGGTTTTCCTCGCGTCGCC ORFCCTGGTGGCCATCTGGGAGTTAAAGAAGGACGTGTACGTGGTGGAGCTGGATTGGTACCCCGACGCCCCGGGCGAGATGGTCGTGCTCACCTGCGATACCCCCGAGGAGGACGGGATCACCTGGACCCTGGACCAATCCAGCGAGGTGCTGGGCAGCGGCAAGACCCTGACCATACAGGTGAAGGAATTTGGGGACGCCGGGCAGTACACCTGCCACAAGGGCGGGGAAGTGCTGTCCCACTCCCTCCTGCTGCTGCATAAGAAGGAGGACGGCATCTGGAGCACCGACATCCTGAAGGACCAAAAGGAGCCCAAGAACAAGACCTTCCTGAGGTGCGAGGCCAAAAACTATTCCGGCCGCTTTACCTGTTGGTGGCTGACCACCATCTCCACCGATCTGACCTTCAGCGTGAAGTCGTCTAGGGGCTCCTCCGACCCCCAGGGCGTAACCTGCGGCGCCGCGACCCTGAGCGCCGAGAGGGTGCGGGGCGATAACAAAGAGTACGAGTACTCGGTGGAGTGCCAGGAGGACAGCGCCTGTCCGGCGGCCGAGGAGAGCCTGCCCATCGAGGTGATGGTGGACGCCGTCCACAAGCTGAAGTACGAGAACTACACCAGTTCGTTCTTCATCAGGGACATCATCAAGCCGGACCCCCCCAAGAACCTCCAGCTGAAGCCCCTGAAGAACAGCAGGCAGGTGGAAGTGTCCTGGGAGTATCCCGACACCTGGAGCACCCCCCACAGCTACTTCAGCCTGACCTTTTGCGTGCAGGTGCAGGGCAAAAGCAAGAGGGAAAAGAAGGACCGGGTGTTCACCGATAAGACGAGCGCCACCGTTATCTGCAGGAAGAACGCCTCCATAAGCGTGAGGGCGCAGGACCGTTACTACAGCAGCAGCTGGAGTGAGTGGGCAAGCGTGCCCTGTAGCGGCGGGGGCGGGGGCGGGTCCCGCAACCTCCCCGTCGCCACCCCCGACCCAGGCATGTTTCCGTGCCTGCACCACAGCCAGAACCTGCTGCGGGCCGTTAGCAACATGCTGCAGAAGGCCAGGCAGACCCTCGAGTTCTATCCCTGCACATCTGAGGAGATCGACCACGAAGACATCACTAAGGATAAGACCTCCACCGTGGAGGCCTGTCTGCCCCTCGAGCTGACCAAGAATGAATCCTGCCTGAACAGCCGAGAGACCAGCTTTATCACCAACGGCTCCTGCCTGGCCAGCAGGAAGACCTCCTTCATGATGGCCCTGTGCCTCTCCAGCATCTACGAGGATCTGAAGATGTACCAGGTAGAGTTCAAGACGATGAACGCCAAGCTCCTGATGGACCCCAAGAGGCAGATATTCCTGGACCAGAACATGCTGGCGGTGATCGACGAGCTGATGCAGGCCCTGAATTTCAACAGCGAGACGGTGCCACAGAAGTCCAGCCTGGAGGAGCCAGACTTCTACAAGACCAAGATCAAACTGTGCATCCTCCTGCACGCGTTCAGGATCCGCGCCGTCACCATAGACAGGGTGATGAGTTATCTGAACGCCAGC  40 hIL12AB_036ATGTGCCATCAGCAGCTGGTAATCAGCTGGTTTAGCCTGGTGTTCCTGGCCAGCCC ORFACTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAACTGGACTGGTACCCCGACGCCCCTGGCGAGATGGTGGTACTGACCTGTGACACCCCGGAGGAAGACGGTATCACCTGGACCCTGGATCAGAGCTCCGAGGTGCTGGGCTCCGGCAAGACACTGACCATCCAAGTTAAGGAATTTGGGGACGCCGGCCAGTACACCTGCCACAAGGGGGGCGAGGTGCTGTCCCACTCCCTGCTGCTTCTGCATAAGAAGGAGGATGGCATCTGGTCCACCGACATACTGAAGGACCAGAAGGAGCCCAAGAATAAGACCTTCCTGAGATGCGAGGCCAAGAACTACTCGGGAAGGTTCACCTGCTGGTGGCTGACCACCATCAGCACCGACCTGACCTTCTCCGTGAAGAGCTCCCGGGGCAGCTCCGACCCCCAGGGCGTAACCTGTGGGGCCGCTACCCTGTCCGCCGAGAGGGTCCGGGGCGACAACAAGGAATACGAGTACAGCGTGGAGTGCCAGGAGGACTCCGCCTGCCCCGCCGCCGAGGAGTCGCTGCCCATAGAGGTGATGGTGGACGCCGTGCACAAGCTCAAGTACGAGAATTACACCAGCAGCTTCTTTATCAGGGACATAATTAAGCCGGACCCCCCAAAGAATCTGCAGCTGAAGCCCCTGAAGAATAGCCGGCAGGTGGAAGTGTCCTGGGAGTACCCCGACACCTGGAGCACCCCCCACTCCTATTTCTCACTGACATTCTGCGTGCAGGTGCAAGGGAAAAGCAAGAGGGAGAAGAAGGATAGGGTGTTCACCGACAAGACAAGCGCCACCGTGATCTGCCGAAAAAATGCCAGCATCAGCGTGAGGGCCCAGGATCGGTATTACAGCAGCTCCTGGAGCGAGTGGGCCAGCGTGCCCTGTTCCGGCGGGGGAGGGGGCGGCTCCCGGAACCTGCCGGTGGCCACCCCCGACCCTGGCATGTTCCCCTGCCTGCATCACAGCCAGAACCTGCTCCGGGCCGTGTCGAACATGCTGCAGAAGGCCCGGCAGACCCTCGAGTTTTACCCCTGCACCAGCGAAGAGATCGACCACGAAGACATAACCAAGGACAAGACCAGCACGGTGGAGGCCTGCCTGCCCCTGGAGCTTACCAAAAACGAGTCCTGCCTGAACAGCCGGGAAACCAGCTTCATAACGAACGGGAGCTGCCTGGCCTCCAGGAAGACCAGCTTCATGATGGCGCTGTGTCTGTCCAGCATATACGAGGATCTGAAGATGTATCAGGTGGAATTCAAAACTATGAATGCCAAGCTCCTGATGGACCCCAAGAGGCAGATCTTCCTGGACCAGAACATGCTAGCCGTGATCGACGAGCTGATGCAGGCCCTCAACTTCAACTCGGAGACGGTGCCCCAGAAGTCCAGCCTCGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATACTGCTGCATGCCTTCAGGATAAGGGCGGTGACTATCGACAGGGTCATGTCCTACCTGAACGCCAGC  41 hIL12AB_037ATGTGCCACCAACAACTGGTGATCAGCTGGTTCTCCCTGGTGTTCCTGGCCAGCCC ORFCCTGGTGGCCATCTGGGAGCTCAAAAAAGACGTGTACGTGGTGGAGCTCGATTGGTACCCAGACGCGCCGGGGGAAATGGTGGTGCTGACCTGCGACACCCCAGAGGAGGATGGCATCACGTGGACGCTGGATCAGTCCAGCGAGGTGCTGGGGAGCGGCAAGACGCTCACCATCCAGGTGAAGGAATTTGGCGACGCGGGCCAGTATACCTGTCACAAGGGCGGCGAGGTGCTGAGCCACTCCCTGCTGCTGCTGCACAAGAAGGAGGATGGGATCTGGTCAACCGATATCCTGAAAGACCAGAAGGAGCCCAAGAACAAGACCTTCCTGCGCTGCGAGGCCAAGAACTATAGCGGCAGGTTCACCTGCTGGTGGCTGACCACCATCAGCACCGACCTGACCTTCAGCGTGAAATCCTCCAGGGGCAGCAGCGACCCCCAGGGCGTGACCTGCGGTGCCGCCACGCTCTCCGCCGAGCGAGTGAGGGGTGACAACAAGGAGTACGAGTACAGCGTGGAATGTCAGGAGGACAGCGCCTGTCCCGCCGCCGAGGAGTCGCTGCCCATCGAGGTGATGGTCGACGCGGTGCACAAGCTCAAATACGAGAATTACACCAGCAGCTTCTTCATCAGGGACATCATCAAGCCCGACCCCCCCAAGAACCTGCAGCTGAAGCCCTTGAAGAACAGCAGGCAGGTGGAGGTGAGCTGGGAGTACCCGGACACCTGGAGCACCCCCCACTCCTACTTCAGCCTGACGTTCTGTGTGCAGGTGCAGGGGAAGTCCAAGAGGGAGAAGAAGGACCGGGTGTTCACCGACAAGACCAGCGCCACCGTGATATGCCGCAAGAACGCGTCCATCAGCGTTCGCGCCCAGGACCGCTACTACAGCAGCTCCTGGTCCGAATGGGCCAGCGTGCCCTGCAGCGGTGGAGGGGGCGGGGGCTCCAGGAATCTGCCGGTGGCCACCCCCGACCCCGGGATGTTCCCGTGTCTGCATCACTCCCAGAACCTGCTGCGGGCCGTGAGCAATATGCTGCAGAAGGCCAGGCAGACGCTCGAGTTCTACCCCTGCACCTCCGAAGAGATCGACCATGAGGACATCACCAAGGACAAGACCAGCACCGTGGAGGCCTGCCTCCCCCTGGAGCTGACCAAAAACGAGAGCTGCCTGAACTCCAGGGAGACCAGCTTTATAACCAACGGCAGCTGCCTCGCCTCCAGGAAGACCTCGTTTATGATGGCCCTCTGCCTGTCCAGCATCTACGAGGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAACGCGAAGTTGCTCATGGACCCCAAGAGGCAGATCTTCCTGGACCAGAACATGCTCGCGGTGATCGACGAGCTGATGCAAGCCCTGAACTTCAACAGCGAGACCGTGCCCCAGAAGAGCAGCCTGGAAGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATCCTGCTGCACGCCTTCCGGATCCGGGCCGTGACCATCGACAGGGTGATGAGCTACCTCAACGCCTCC  42 hIL12AB_038ATGTGCCACCAGCAGCTCGTGATCAGCTGGTTCTCCCTCGTCTTCCTGGCCTCCCC ORFGCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGCTGGACTGGTATCCCGACGCCCCCGGCGAGATGGTGGTGCTGACGTGCGACACACCAGAAGAGGACGGGATCACATGGACCCTGGATCAGTCGTCCGAGGTGCTGGGGAGCGGCAAGACCCTCACCATCCAAGTGAAGGAGTTCGGGGACGCCGGCCAGTACACCTGCCACAAGGGCGGGGAGGTGCTCTCCCATAGCCTGCTCCTCCTGCACAAAAAGGAGGATGGCATCTGGAGCACCGACATCCTGAAGGACCAGAAGGAGCCCAAGAACAAGACATTTCTCAGGTGTGAGGCCAAGAACTATTCGGGCAGGTTTACCTGTTGGTGGCTCACCACCATCTCTACCGACCTGACGTTCTCCGTCAAGTCAAGCAGGGGGAGCTCGGACCCCCAGGGGGTGACATGTGGGGCCGCCACCCTGAGCGCGGAGCGTGTCCGCGGCGACAACAAGGAGTACGAGTATTCCGTGGAGTGCCAGGAGGACAGCGCCTGCCCCGCCGCCGAGGAGTCCCTGCCCATAGAGGTGATGGTGGACGCCGTCCACAAGTTGAAGTACGAAAATTATACCTCCTCGTTCTTCATTAGGGACATCATCAAGCCTGACCCCCCGAAGAACCTACAACTCAAGCCCCTCAAGAACTCCCGCCAGGTGGAGGTGTCCTGGGAGTACCCCGACACCTGGTCCACCCCGCACAGCTACTTCAGCCTGACCTTCTGCGTGCAGGTCCAGGGGAAGAGCAAGCGTGAAAAGAAAGACAGGGTGTTCACCGACAAGACGAGCGCCACCGTGATCTGCAGGAAAAACGCCTCCATCTCCGTGCGCGCCCAGGACAGGTACTACAGTAGCTCCTGGAGCGAATGGGCCAGCGTGCCGTGCAGCGGCGGGGGAGGAGGCGGCAGTCGCAACCTGCCCGTGGCCACCCCCGACCCCGGCATGTTCCCATGCCTGCACCACAGCCAGAACCTGCTGAGGGCAGTCAGCAATATGCTGCAGAAGGCCAGGCAGACCCTGGAGTTTTATCCCTGCACCAGCGAGGAGATCGACCACGAGGACATCACCAAGGACAAGACCTCCACCGTCGAGGCCTGCCTGCCACTGGAGCTGACCAAAAACGAGAGCTGCCTGAACTCCAGGGAGACCTCCTTCATCACCAACGGGAGCTGCCTGGCCAGCCGGAAGACCAGCTTCATGATGGCGCTGTGCCTCAGCAGCATCTACGAGGATCTCAAGATGTACCAGGTGGAGTTCAAGACCATGAACGCGAAGCTGCTGATGGACCCCAAGCGGCAGATCTTCCTGGACCAGAACATGCTGGCCGTGATTGACGAGCTCATGCAGGCCCTGAACTTCAATAGCGAGACCGTCCCCCAAAAGAGCAGCCTGGAGGAACCCGACTTCTACAAAACGAAGATCAAGCTCTGCATCCTGCTGCACGCCTTCCGGATCCGGGCCGTGACCATCGATCGTGTGATGAGCTACCTGAACGCCTCG  43 hIL12AB_039ATGTGCCACCAGCAGCTCGTCATCTCCTGGTTTAGCCTGGTGTTTCTGGCCTCCCC ORFCCTGGTCGCCATCTGGGAGCTGAAGAAAGACGTGTACGTGGTGGAGCTGGACTGGTACCCGGACGCTCCCGGGGAGATGGTGGTGCTGACCTGCGACACCCCCGAGGAGGACGGCATCACCTGGACCCTGGACCAGAGCTCCGAGGTGCTGGGGAGCGGCAAGACCCTGACCATTCAGGTGAAAGAGTTCGGCGACGCCGGCCAATATACCTGCCACAAGGGGGGGGAGGTCCTGTCGCATTCCCTGCTGCTGCTTCACAAAAAGGAGGATGGCATCTGGAGCACCGACATCCTGAAGGACCAGAAAGAACCCAAGAACAAGACGTTCCTGCGCTGCGAGGCCAAGAACTACAGCGGCCGGTTCACCTGTTGGTGGCTGACCACCATCTCCACCGACCTGACTTTCTCGGTGAAGAGCAGCCGCGGGAGCAGCGACCCCCAGGGAGTGACCTGCGGCGCCGCCACCCTGAGCGCCGAAAGGGTGAGGGGCGACAATAAAGAGTACGAGTATTCCGTGGAGTGCCAGGAGGACAGCGCCTGTCCCGCCGCCGAGGAGTCCCTGCCTATCGAGGTGATGGTCGACGCGGTGCACAAGCTCAAGTACGAAAACTACACCAGCAGCTTTTTCATCAGGGATATCATCAAACCAGACCCCCCCAAGAACCTGCAGCTGAAGCCCCTGAAAAACAGCAGGCAGGTGGAAGTGAGCTGGGAATACCCCGATACCTGGTCCACCCCCCACAGCTACTTCAGCCTGACCTTCTGCGTGCAGGTGCAGGGGAAGTCCAAGCGGGAGAAGAAAGATCGGGTGTTCACGGACAAGACCAGCGCCACCGTGATTTGCAGGAAAAACGCCAGCATCTCCGTGAGGGCTCAGGACAGGTACTACAGCTCCAGCTGGAGCGAGTGGGCCTCCGTGCCTTGCAGCGGGGGAGGAGGCGGCGGCAGCAGGAATCTGCCCGTCGCAACCCCCGACCCCGGCATGTTCCCCTGCCTGCACCACAGCCAGAATCTGCTGCGAGCCGTGAGCAACATGCTCCAGAAGGCCCGGCAGACGCTGGAGTTCTACCCCTGCACCTCCGAGGAGATCGACCACGAGGACATCACCAAGGATAAGACGAGCACCGTCGAGGCCTGTCTCCCCCTGGAGCTCACCAAGAACGAGTCCTGCCTGAATAGCAGGGAGACGTCCTTCATAACCAACGGCAGCTGTCTGGCGTCCAGGAAGACCAGCTTCATGATGGCCCTCTGCCTGAGCTCCATCTACGAGGACCTCAAGATGTACCAGGTCGAGTTCAAGACCATGAACGCAAAACTGCTCATGGATCCAAAGAGGCAGATCTTTCTGGACCAGAACATGCTGGCCGTGATCGATGAACTCATGCAGGCCCTGAATTTCAATTCCGAGACCGTGCCCCAGAAGAGCTCCCTGGAGGAACCCGACTTCTACAAAACAAAGATCAAGCTGTGTATCCTCCTGCACGCCTTCCGGATCAGGGCCGTCACCATTGACCGGGTGATGTCCTACCTGAACGCCAGC  44 hIL12AB_040ATGTGCCATCAGCAGCTGGTGATCAGCTGGTTCAGCCTCGTGTTCCTCGCCAGCCC ORFCCTCGTGGCCATCTGGGAGCTGAAAAAGGACGTGTACGTGGTGGAGCTGGACTGGTATCCCGACGCCCCGGGCGAGATGGTGGTGCTGACCTGCGACACCCCCGAGGAGGACGGCATTACCTGGACACTGGACCAGAGCAGCGAGGTCCTGGGCAGCGGGAAGACCCTGACAATTCAGGTGAAGGAGTTCGGCGACGCCGGACAGTACACGTGCCACAAGGGGGGGGAGGTGCTGTCCCACAGCCTCCTCCTGCTGCACAAGAAGGAGGATGGCATCTGGAGCACCGACATCCTGAAGGATCAGAAGGAGCCCAAGAACAAGACCTTTCTGAGATGCGAGGCCAAGAATTACAGCGGCCGTTTCACCTGCTGGTGGCTCACCACCATCAGCACCGACCTGACCTTCAGCGTGAAATCCTCCAGGGGCTCCTCCGACCCGCAGGGAGTGACCTGCGGCGCCGCCACACTGAGCGCCGAGCGGGTCAGAGGGGACAACAAGGAGTACGAGTACAGCGTTGAGTGCCAGGAGGACAGCGCCTGTCCCGCGGCCGAGGAATCCCTGCCCATCGAGGTGATGGTGGACGCAGTGCACAAGCTGAAGTACGAGAACTATACCTCGAGCTTCTTCATCCGGGATATCATTAAGCCCGATCCCCCGAAGAACCTGCAGCTCAAACCCCTGAAGAACAGCAGGCAGGTGGAGGTCTCCTGGGAGTACCCCGACACATGGTCCACCCCCCATTCCTATTTCTCCCTGACCTTTTGCGTGCAGGTGCAGGGCAAGAGCAAGAGGGAGAAAAAGGACAGGGTGTTCACCGACAAGACCTCCGCCACCGTGATCTGCCGTAAGAACGCTAGCATCAGCGTCAGGGCCCAGGACAGGTACTATAGCAGCTCCTGGTCCGAGTGGGCCAGCGTCCCGTGCAGCGGCGGGGGCGGTGGAGGCTCCCGGAACCTCCCCGTGGCCACCCCGGACCCCGGGATGTTTCCCTGCCTGCATCACAGCCAGAACCTGCTGAGGGCCGTGTCCAACATGCTGCAGAAGGCCAGGCAGACACTCGAGTTTTACCCCTGCACCAGCGAGGAGATCGACCACGAAGACATCACCAAGGACAAGACCTCCACCGTGGAGGCATGCCTGCCCCTGGAGCTGACCAAAAACGAAAGCTGTCTGAACTCCAGGGAGACCTCCTTTATCACGAACGGCTCATGCCTGGCCTCCAGAAAGACCAGCTTCATGATGGCCCTGTGCCTGAGCTCCATCTACGAGGACTTGAAAATGTACCAGGTCGAGTTCAAGACCATGAACGCCAAGCTGCTCATGGACCCCAAAAGGCAGATCTTTCTGGACCAGAATATGCTGGCCGTGATCGACGAGCTCATGCAAGCCCTGAATTTCAACAGCGAGACCGTGCCCCAGAAGTCCTCCCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATACTCCTGCACGCGTTTAGGATCAGGGCGGTGACCATCGATAGGGTGATGAGCTACCTGAATGCCTCC  45 Wild Type MCHQQLVISWFSLVFLASPLVAIL12B signal peptide Amino acids  46 Wild TypeATGTGTCACCAGCAGTTGGTCATCTCTTGGTTTTCCCTGGTTTTTCTGGCATCTCC IL12B signalCCTCGTGGCC peptide Nucleic acids  47 Syn 5 ATTGGGCACCCGTAAGGG promoter 48 Signal peptide-MCHQQLVISWFSLVFLASPLVAIWELKKDVYVVELDWYPDAPGEMVVLTCDTPEED IL12B-linker-GITWTLDQSSEVLGSGKTLTIQVKEFGDAGQYTCHKGGEVLSHSLLLLHKKEDGIW IL12A aminoSTDILKDQKEPKNKTFLRCEAKNYSGRFTCWWLTTISTDLTFSVKSSRGSSDPQGV acid sequenceTCGAATLSAERVRGDNKEYEYSVECQEDSACPAAEESLPIEVMVDAVHKLKYENYT #1SSFFIRDIIKPDPPKNLQLKPLKNSRQVEVSWEYPDTWSTPHSYFSLTFCVQVQGKSKREKKDRVFTDKTSATVICRKNASISVRAQDRYYSSSWSEWASVPCSGGGGGGSRNLPVATPDPGMFPCLHHSQNLLRAVSNMLQKARQTLEFYPCTSEEIDHEDITKDKTSTVEACLPLELTKNESCLNSRETSFITNGSCLASRKTSFMMALCLSSIYEDLKMYQVEFKTMNAKLLMDPKRQIFLDQNMLAVIDELMQALNFNSETVPQKSSLEEPDFYKTKIKLCILLHAFRIRAVTIDRVMSYLNAS  49 Signal peptide-MGCHQQLVISWFSLVFLASPLVAIWELKKDVYVVELDWYPDAPGEMVVLTCDTPEE IL12B-linker-DGITWTLDQSSEVLGSGKTLTIQVKEFGDAGQYTCHKGGEVLSHSLLLLHKKEDGI IL12A aminoWSTDILKDQKEPKNKTFLRCEAKNYSGRFTCWWLTTISTDLTFSVKSSRGSSDPQG acid sequenceVTCGAATLSAERVRGDNKEYEYSVECQEDSACPAAEESLPIEVMVDAVHKLKYENY #2TSSFFIRDIIKPDPPKNLQLKPLKNSRQVEVSWEYPDTWSTPHSYFSLTFCVQVQGKSKREKKDRVFTDKTSATVICRKNASISVRAQDRYYSSSWSEWASVPCSGGGGGGSRNLPVATPDPGMFPCLHHSQNLLRAVSNMLQKARQTLEFYPCTSEEIDHEDITKDKTSTVEACLPLELTKNESCLNSRETSFITNGSCLASRKTSFMMALCLSSIYEDLKMYQVEFKTMNAKLLMDPKRQIFLDQNMLAVIDELMQALNFNSETVPQKSSLEEPDFYKTKIKLCILLHAFRIRAVTIDRVMSYLNAS  50 miR-122CCUUAGCAGAGCUGUGGAGUGUGACAAUGGUGUUUGUGUCUAAACUAUCAAACGCCAUUAUCACACUAAAUAGCUACUGCUAGGC  51 miR-122-3p AACGCCAUUAUCACACUAAAUA  52miR-122-3p UAUUUAGUGUGAUAAUGGCGUU binding site  53 miR-122-5pUGGAGUGUGAGAAUGGUGUUUG  54 miR-122-5p CAAACACCAUUGUCACACUCCAbinding site  55 5′UTR-017GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUUUAAGAGCCACC  56 5′UTR-018UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACC  57 142-3p UGAUAAUAGUCCAUAAAGUAGGAAACACUACAGCUGGAGCCUCGGUGGCCAUGCUU 5′UTR-001CUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC  58 142-3p UGAUAAUAGGCUGGAGCCUCGGUGGCUCCAUAAAGUAGGAAACACUACACAUGCUU 5′UTR-002CUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC  59 142-3pUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUCCAUAAAGUAGGA 5′UTR-003AACACUACAUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC  60 142-3pUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCA 5′UTR-004GUCCAUAAAGUAGGAAACACUACACCCCUCCUCCCCUUCCUGCACCCGUACCCCCGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC  61 142-3pUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCA 5′UTR-005GCCCCUCCUCCCCUUCUCCAUAAAGUAGGAAACACUACACUGCACCCGUACCCCCGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC  62 142-3pUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCA 5′UTR-006GCCCCUCCUCCCCUUCCUGCACCCGUACCCCCUCCAUAAAGUAGGAAACACUACAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC  63 142-3pUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCA 5′UTR-007GCCCCUCCUCCCCUUCCUGCACCCGUACCCCCGUGGUCUUUGAAUAAAGUUCCAUAAAGUAGGAAACACUACACUGAGUGGGCGGC  64 3′UTR-001GCGCCUGCCCACCUGCCACCGACUGCUGGAACCCAGCCAGUGGGAGGGCCUGGCCC (CreatineACCAGAGUCCUGCUCCCUCACUCCUCGCCCCGCCCCCUGUCCCAGAGUCCCACCUG KinaseGGGGCUCUCUCCACCCUUCUCAGAGUUCCAGUUUCAACCAGAGUUCCAACCAAUGG UTR)GCUCCAUCCUCUGGAUUCUGGCCAAUGAAAUAUCUCCCUGGCAGGGUCCUCUUCUUUUCCCAGAGCUCCACCCCAACCAGGAGCUCUAGUUAAUGGAGAGCUCCCAGCACACUCGGAGCUUGUGCUUUGUCUCCACGCAAAGCGAUAAAUAAAAGCAUUGGUGGCCUUUGGUCUUUGAAUAAAGCCUGAGUAGGAAGUCUAGA  65 3′UTR-002GCCCCUGCCGCUCCCACCCCCACCCAUCUGGGCCCCGGGUUCAAGAGAGAGCGGGG (MyoglobinUCUGAUCUCGUGUAGCCAUAUAGAGUUUGCUUCUGAGUGUCUGCUUUGUUUAGUAG UTR)AGGUGGGCAGGAGGAGCUGAGGGGCUGGGGCUGGGGUGUUGAAGUUGGCUUUGCAUGCCCAGCGAUGCGCCUCCCUGUGGGAUGUCAUCACCCUGGGAACCGGGAGUGGCCCUUGGCUCACUGUGUUCUGCAUGGUUUGGAUCUGAAUUAAUUGUCCUUUCUUCUAAAUCCCAACCGAACUUCUUCCAACCUCCAAACUGGCUGUAACCCCAAAUCCAAGCCAUUAACUACACCUGACAGUAGCAAUUGUCUGAUUAAUCACUGGCCCCUUGAAGACAGCAGAAUGUCCCUUUGCAAUGAGGAGGAGAUCUGGGCUGGGCGGGCCAGCUGGGGAAGCAUUUGACUAUCUGGAACUUGUGUGUGCCUCCUCAGGUAUGGCAGUGACUCACCUGGUUUUAAUAAAACAACCUGCAACAUCUCAUGGUCUUUGAAUAAAGCCUGAGUAGGA AGUCUAGA  663′UTR-003 ACACACUCCACCUCCAGCACGCGACUUCUCAGGACGACGAAUCUUCUCAAUGGGGG(α-actin GGCGGCUGAGCUCCAGCCACCCCGCAGUCACUUUCUUUGUAACAACUUCCGUUGCU UTR)GCCAUCGUAAACUGACACAGUGUUUAUAACGUGUACAUACAUUAACUUAUUACCUCAUUUUGUUAUUUUUCGAAACAAAGCCCUGUGGAAGAAAAUGGAAAACUUGAAGAAGCAUUAAAGUCAUUCUGUUAAGCUGCGUAAAUGGUCUUUGAAUAAAGCCUGAGUAGG AAGUCUAGA  673′UTR-004 CAUCACAUUUAAAAGCAUCUCAGCCUACCAUGAGAAUAAGAGAAAGAAAAUGAAGA(Albumin UCAAAAGCUUAUUCAUCUGUUUUUCUUUUUCGUUGGUGUAAAGCCAACACCCUGUC UTR)UAAAAAACAUAAAUUUCUUUAAUCAUUUUGCCUCUUUUCUCUGUGCUUCAAUUAAUAAAAAAUGGAAAGAAUCUAAUAGAGUGGUACAGCACUGUUAUUUUUCAAAGAUGUGUUGCUAUCCUGAAAAUUCUGUAGGUUCUGUGGAAGUUCCAGUGUUCUCUCUUAUUCCACUUCGGUAGAGGAUUUCUAGUUUCUUGUGGGCUAAUUAAAUAAAUCAUUAAUACUCUUCUAAUGGUCUUUGAAUAAAGCCUGAGUAGGAAGUCUAGA  68 3′UTR-005 (α-GCUGCCUUCUGCGGGGCUUGCCUUCUGGCCAUGCCCUUCUUCUCUCCCUUGCACCU globin UTR)GUACCUCUUGGUCUUUGAAUAAAGCCUGAGUAGGAAGGCGGCCGCUCGAGCAUGCA UCUAGA  693′UTR-006 GCCAAGCCCUCCCCAUCCCAUGUAUUUAUCUCUAUUUAAUAUUUAUGUCUAUUUAA(G-CSF UTR) GCCUCAUAUUUAAAGACAGGGAAGAGCAGAACGGAGCCCCAGGCCUCUGUGUCCUUCCCUGCAUUUCUGAGUUUCAUUCUCCUGCCUGUAGCAGUGAGAAAAAGCUCCUGUCCUCCCAUCCCCUGGACUGGGAGGUAGAUAGGUAAAUACCAAGUAUUUAUUACUAUGACUGCUCCCCAGCCCUGGCUCUGCAAUGGGCACUGGGAUGAGCCGCUGUGAGCCCCUGGUCCUGAGGGUCCCCACCUGGGACCCUUGAGAGUAUCAGGUCUCCCACGUGGGAGACAAGAAAUCCCUGUUUAAUAUUUAAACAGCAGUGUUCCCCAUCUGGGUCCUUGCACCCCUCACUCUGGCCUCAGCCGACUGCACAGCGGCCCCUGCAUCCCCUUGGCUGUGAGGCCCCUGGACAAGCAGAGGUGGCCAGAGCUGGGAGGCAUGGCCCUGGGGUCCCACGAAUUUGCUGGGGAAUCUCGUUUUUCUUCUUAAGACUUUUGGGACAUGGUUUGACUCCCGAACAUCACCGACGCGUCUCCUGUUUUUCUGGGUGGCCUCGGGACACCUGCCCUGCCCCCACGAGGGUCAGGACUGUGACUCUUUUUAGGGCCAGGCAGGUGCCUGGACAUUUGCCUUGCUGGACGGGGACUGGGGAUGUGGGAGGGAGCAGACAGGAGGAAUCAUGUCAGGCCUGUGUGUGAAAGGAAGCUCCACUGUCACCCUCCACCUCUUCACCCCCCACUCACCAGUGUCCCCUCCACUGUCACAUUGUAACUGAACUUCAGGAUAAUAAAGUGUUUGCCUCCAUGGUCUUUGAAUAAAGCCUGAGUAGGAAGGCGGCCGCUCGAG CAUGCAUCUAGA 70 3′UTR-007 ACUCAAUCUAAAUUAAAAAAGAAAGAAAUUUGAAAAAACUUUCUCUUUGCCAUUUC(Colla2; UUCUUCUUCUUUUUUAACUGAAAGCUGAAUCCUUCCAUUUCUUCUGCACAUCUACUcollagen, Type UGCUUAAAUUGUGGGCAAAAGAGAAAAAGAAGGAUUGAUCAGAGCAUUGUGCAAUAI, alpha 2 CAGUUUCAUUAACUCCUUCCCCCGCUCCCCCAAAAAUUUGAAUUUUUUUUUCAACA UTR)CUCUUACACCUGUUAUGGAAAAUGUCAACCUUUGUAAGAAAACCAAAAUAAAAAUUGAAAAAUAAAAACCAUAAACAUUUGCACCACUUGUGGCUUUUGAAUAUCUUCCACAGAGGGAAGUUUAAAACCCAAACUUCCAAAGGUUUAAACUACCUCAAAACACUUUCCCAUGAGUGUGAUCCACAUUGUUAGGUGCUGACCUAGACAGAGAUGAACUGAGGUCCUUGUUUUGUUUUGUUCAUAAUACAAAGGUGCUAAUUAAUAGUAUUUCAGAUACUUGAAGAAUGUUGAUGGUGCUAGAAGAAUUUGAGAAGAAAUACUCCUGUAUUGAGUUGUAUCGUGUGGUGUAUUUUUUAAAAAAUUUGAUUUAGCAUUCAUAUUUUCCAUCUUAUUCCCAAUUAAAAGUAUGCAGAUUAUUUGCCCAAAUCUUCUUCAGAUUCAGCAUUUGUUCUUUGCCAGUCUCAUUUUCAUCUUCUUCCAUGGUUCCACAGAAGCUUUGUUUCUUGGGCAAGCAGAAAAAUUAAAUUGUACCUAUUUUGUAUAUGUGAGAUGUUUAAAUAAAUUGUGAAAAAAAUGAAAUAAAGCAUGUUUGGUUUUCCAAAAGAACAUAU  71 3′UTR-008CGCCGCCGCCCGGGCCCCGCAGUCGAGGGUCGUGAGCCCACCCCGUCCAUGGUGCU (Col6a2;AAGCGGGCCCGGGUCCCACACGGCCAGCACCGCUGCUCACUCGGACGACGCCCUGG collagen, TypeGCCUGCACCUCUCCAGCUCCUCCCACGGGGUCCCCGUAGCCCCGGCCCCCGCCCAG VI, alpha 2CCCCAGGUCUCCCCAGGCCCUCCGCAGGCUGCCCGGCCUCCCUCCCCCUGCAGCCA UTR)UCCCAAGGCUCCUGACCUACCUGGCCCCUGAGCUCUGGAGCAAGCCCUGACCCAAUAAAGGCUUUGAACCCAU  72 3′UTR-009GGGGCUAGAGCCCUCUCCGCACAGCGUGGAGACGGGGCAAGGAGGGGGGUUAUUAG (RPN1;GAUUGGUGGUUUUGUUUUGCUUUGUUUAAAGCCGUGGGAAAAUGGCACAACUUUAC ribophorin ICUCUGUGGGAGAUGCAACACUGAGAGCCAAGGGGUGGGAGUUGGGAUAAUUUUUAU UTR)AUAAAAGAAGUUUUUCCACUUUGAAUUGGUAAAAGUGGCAUUUUUCCUAUGUGCAGUCACUCCUCUCAUUUCUAAAAUAGGGACGUGGCCAGGCACGGUGGCUCAUGCCUGUAAUCCCAGCACUUUGGGAGGCCGAGGCAGGCGGCUCACGAGGUCAGGAGAUCGAGACUAUCCUGGCUAACACGGUAAAACCCUGUCUCUACUAAAAGUACAAAAAAUUAGCUGGGCGUGGUGGUGGGCACCUGUAGUCCCAGCUACUCGGGAGGCUGAGGCAGGAGAAAGGCAUGAAUCCAAGAGGCAGAGCUUGCAGUGAGCUGAGAUCACGCCAUUGCACUCCAGCCUGGGCAACAGUGUUAAGACUCUGUCUCAAAUAUAAAUAAAUAAAUAAAUAAAUAAAUAAAUAAAUAAAAAUAAAGCGAGAUGUUGCCCUCAAA  73 3′UTR-010GGCCCUGCCCCGUCGGACUGCCCCCAGAAAGCCUCCUGCCCCCUGCCAGUGAAGUC (LRP1; lowCUUCAGUGAGCCCCUCCCCAGCCAGCCCUUCCCUGGCCCCGCCGGAUGUAUAAAUG densityUAAAAAUGAAGGAAUUACAUUUUAUAUGUGAGCGAGCAAGCCGGCAAGCGAGCACA lipoproteinGUAUUAUUUCUCCAUCCCCUCCCUGCCUGCUCCUUGGCACCCCCAUGCUGCCUUCA receptor-GGGAGACAGGCAGGGAGGGCUUGGGGCUGCACCUCCUACCCUCCCACCAGAACGCA related proteinCCCCACUGGGAGAGCUGGUGGUGCAGCCUUCCCCUCCCUGUAUAAGACACUUUGCC 1 UTR)AAGGCUCUCCCCUCUCGCCCCAUCCCUGCUUGCCCGCUCCCACAGCUUCCUGAGGGCUAAUUCUGGGAAGGGAGAGUUCUUUGCUGCCCCUGUCUGGAAGACGUGGCUCUGGGUGAGGUAGGCGGGAAAGGAUGGAGUGUUUUAGUUCUUGGGGGAGGCCACCCCAAACCCCAGCCCCAACUCCAGGGGCACCUAUGAGAUGGCCAUGCUCAACCCCCCUCCCAGACAGGCCCUCCCUGUCUCCAGGGCCCCCACCGAGGUUCCCAGGGCUGGAGACUUCCUCUGGUAAACAUUCCUCCAGCCUCCCCUCCCCUGGGGACGCCAAGGAGGUGGGCCACACCCAGGAAGGGAAAGCGGGCAGCCCCGUUUUGGGGACGUGAACGUUUUAAUAAUUUUUGCUGAAUUCCUUUACAACUAAAUAACACAGAUAUUGUUAUAAAUAAAAUUGU  74 3′UTR-011AUAUUAAGGAUCAAGCUGUUAGCUAAUAAUGCCACCUCUGCAGUUUUGGGAACAGG (Nnt1;CAAAUAAAGUAUCAGUAUACAUGGUGAUGUACAUCUGUAGCAAAGCUCUUGGAGAA cardiotrophin-AAUGAAGACUGAAGAAAGCAAAGCAAAAACUGUAUAGAGAGAUUUUUCAAAAGCAG like cytokineUAAUCCCUCAAUUUUAAAAAAGGAUUGAAAAUUCUAAAUGUCUUUCUGUGCAUAUU factor 1 UTR)UUUUGUGUUAGGAAUCAAAAGUAUUUUAUAAAAGGAGAAAGAACAGCCUCAUUUUAGAUGUAGUCCUGUUGGAUUUUUUAUGCCUCCUCAGUAACCAGAAAUGUUUUAAAAAACUAAGUGUUUAGGAUUUCAAGACAACAUUAUACAUGGCUCUGAAAUAUCUGACACAAUGUAAACAUUGCAGGCACCUGCAUUUUAUGUUUUUUUUUUCAACAAAUGUGACUAAUUUGAAACUUUUAUGAACUUCUGAGCUGUCCCCUUGCAAUUCAACCGCAGUUUGAAUUAAUCAUAUCAAAUCAGUUUUAAUUUUUUAAAUUGUACUUCAGAGUCUAUAUUUCAAGGGCACAUUUUCUCACUACUAUUUUAAUACAUUAAAGGACUAAAUAAUCUUUCAGAGAUGCUGGAAACAAAUCAUUUGCUUUAUAUGUUUCAUUAGAAUACCAAUGAAACAUACAACUUGAAAAUUAGUAAUAGUAUUUUUGAAGAUCCCAUUUCUAAUUGGAGAUCUCUUUAAUUUCGAUCAACUUAUAAUGUGUAGUACUAUAUUAAGUGCACUUGAGUGGAAUUCAACAUUUGACUAAUAAAAUGAGUUCAUCAUGUUGGCAAGUGAUGUGGCAAUUAUCUCUGGUGACAAAAGAGUAAAAUCAAAUAUUUCUGCCUGUUACAAAUAUCAAGGAAGACCUGCUACUAUGAAAUAGAUGACAUUAAUCUGUCUUCACUGUUUAUAAUACGGAUGGAUUUUUUUUCAAAUCAGUGUGUGUUUUGAGGUCUUAUGUAAUUGAUGACAUUUGAGAGAAAUGGUGGCUUUUUUUAGCUACCUCUUUGUUCAUUUAAGCACCAGUAAAGAUCAUGUCUUUUUAUAGAAGUGUAGAUUUUCUUUGUGACUUUGCUAUCGUGCCUAAAGCUCUAAAUAUAGGUGAAUGUGUGAUGAAUACUCAGAUUAUUUGUCUCUCUAUAUAAUUAGUUUGGUACUAAGUUUCUCAAAAAAUUAUUAACACAUGAAAGACAAUCUCUAAACCAGAAAAAGAAGUAGUACAAAUUUUGUUACUGUAAUGCUCGCGUUUAGUGAGUUUAAAACACACAGUAUCUUUUGGUUUUAUAAUCAGUUUCUAUUUUGCUGUGCCUGAGAUUAAGAUCUGUGUAUGUGUGUGUGUGUGUGUGUGCGUUUGUGUGUUAAAGCAGAAAAGACUUUUUUAAAAGUUUUAAGUGAUAAAUGCAAUUUGUUAAUUGAUCUUAGAUCACUAGUAAACUCAGGGCUGAAUUAUACCAUGUAUAUUCUAUUAGAAGAAAGUAAACACCAUCUUUAUUCCUGCCCUUUUUCUUCUCUCAAAGUAGUUGUAGUUAUAUCUAGAAAGAAGCAAUUUUGAUUUCUUGAAAAGGUAGUUCCUGCACUCAGUUUAAACUAAAAAUAAUCAUACUUGGAUUUUAUUUAUUUUUGUCAUAGUAAAAAUUUUAAUUUAUAUAUAUUUUUAUUUAGUAUUAUCUUAUUCUUUGCUAUUUGCCAAUCCUUUGUCAUCAAUUGUGUUAAAUGAAUUGAAAAUUCAUGCCCUGUUCAUUUUAUUUUACUUUAUUGGUUAGGAUAUUUAAAGGAUUUUUGUAUAUAUAAUUUCUUAAAUUAAUAUUCCAAAAGGUUAGUGGACUUAGAUUAUAAAUUAUGGCAAAAAUCUAAAAACAACAAAAAUGAUUUUUAUACAUUCUAUUUCAUUAUUCCUCUUUUUCCAAUAAGUCAUACAAUUGGUAGAUAUGACUUAUUUUAUUUUUGUAUUAUUCACUAUAUCUUUAUGAUAUUUAAGUAUAAAUAAUUAAAAAAAUUUAUUGUACCUUAUAGUCUGUCACCAAAAAAAAAAAAUUAUCUGUAGGUAGUGAAAUGCUAAUGUUGAUUUGUCUUUAAGGGCUUGUUAACUAUCCUUUAUUUUCUCAUUUGUCUUAAAUUAGGAGUUUGUGUUUAAAUUACUCAUCUAAGCAAAAAAUGUAUAUAAAUCCCAUUACUGGGUAUAUACCCAAAGGAUUAUAAAUCAUGCUGCUAUAAAGACACAUGCACACGUAUGUUUAUUGCAGCACUAUUCACAAUAGCAAAGACUUGGAACCAACCCAAAUGUCCAUCAAUGAUAGACUUGAUUAAGAAAAUGUGCACAUAUACACCAUGGAAUACUAUGCAGCCAUAAAAAAGGAUGAGUUCAUGUCCUUUGUAGGGACAUGGAUAAAGCUGGAAACCAUCAUUCUGAGCAAACUAUUGCAAGGACAGAAAACCAAACACUGCAUGUUCUCACUCAUAGGUGGGAAUUGAACAAUGAGAACACUUGGACACAAGGUGGGGAACACCACACACCAGGGCCUGUCAUGGGGUGGGGGGAGUGGGGAGGGAUAGCAUUAGGAGAUAUACCUAAUGUAAAUGAUGAGUUAAUGGGUGCAGCACACCAACAUGGCACAUGUAUACAUAUGUAGCAAACCUGCACGUUGUGCACAUGUACCCUAGAACUUAAAGUAUAAUUAAAAAAAAAAAGAAAACAGAAGCUAUUUAUAAAGAAGUUAUUUGCUGAAAUAAAUGUGAUCUUUCCCAUUAAAAAAAUAAAGAAAUUUUGGGGUAAAAAAACACAAUAUAUUGUAUUCUUGAAAAAUUCUAAGAGAGUGGAUGUGAAGUGUUCUCACCACAAAAGUGAUAACUAAUUGAGGUAAUGCACAUAUUAAUUAGAAAGAUUUUGUCAUUCCACAAUGUAUAUAUACUUAAAAAUAUGUUAUACACAAUAAAUACAUACAUUAAAAAAUAAGUAAAUGUA  75 3′UTR-012CCCACCCUGCACGCCGGCACCAAACCCUGUCCUCCCACCCCUCCCCACUCAUCACU (Col6al;AAACAGAGUAAAAUGUGAUGCGAAUUUUCCCGACCAACCUGAUUCGCUAGAUUUUU collagen, TypeUUUAAGGAAAAGCUUGGAAAGCCAGGACACAACGCUGCUGCCUGCUUUGUGCAGGG VI, alpha 1UCCUCCGGGGCUCAGCCCUGAGUUGGCAUCACCUGCGCAGGGCCCUCUGGGGCUCA UTR)GCCCUGAGCUAGUGUCACCUGCACAGGGCCCUCUGAGGCUCAGCCCUGAGCUGGCGUCACCUGUGCAGGGCCCUCUGGGGCUCAGCCCUGAGCUGGCCUCACCUGGGUUCCCCACCCCGGGCUCUCCUGCCCUGCCCUCCUGCCCGCCCUCCCUCCUGCCUGCGCAGCUCCUUCCCUAGGCACCUCUGUGCUGCAUCCCACCAGCCUGAGCAAGACGCCCUCUCGGGGCCUGUGCCGCACUAGCCUCCCUCUCCUCUGUCCCCAUAGCUGGUUUUUCCCACCAAUCCUCACCUAACAGUUACUUUACAAUUAAACUCAAAGCAAGCUCUUCUCCUCAGCUUGGGGCAGCCAUUGGCCUCUGUCUCGUUUUGGGAAACCAAGGUCAGGAGGCCGUUGCAGACAUAAAUCUCGGCGACUCGGCCCCGUCUCCUGAGGGUCCUGCUGGUGACCGGCCUGGACCUUGGCCCUACAGCCCUGGAGGCCGCUGCUGACCAGCACUGACCCCGACCUCAGAGAGUACUCGCAGGGGCGCUGGCUGCACUCAAGACCCUCGAGAUUAACGGUGCUAACCCCGUCUGCUCCUCCCUCCCGCAGAGACUGGGGCCUGGACUGGACAUGAGAGCCCCUUGGUGCCACAGAGGGCUGUGUCUUACUAGAAACAACGCAAACCUCUCCUUCCUCAGAAUAGUGAUGUGUUCGACGUUUUAUCAAAGGCCCCCUUUCUAUGUUCAUGUUAGUUUUGCUCCUUCUGUGUUUUUUUCUGAACCAUAUCCAUGUUGCUGACUUUUCCAAAUAAAGGUUUUCACUCCUCUC  76 3′UTR-013AGAGGCCUGCCUCCAGGGCUGGACUGAGGCCUGAGCGCUCCUGCCGCAGAGCUGGC (Calr;CGCGCCAAAUAAUGUCUCUGUGAGACUCGAGAACUUUCAUUUUUUUCCAGGCUGGU calreticulinUCGGAUUUGGGGUGGAUUUUGGUUUUGUUCCCCUCCUCCACUCUCCCCCACCCCCU UTR)CCCCGCCCUUUUUUUUUUUUUUUUUUAAACUGGUAUUUUAUCUUUGAUUCUCCUUCAGGCCUCACCCCUGGUUCUCAUCUUUCUUGAUCAACAUCUUUUCUUGCCUCUGUCCCCUUCUCUCAUCUCUUAGCUCCCCUCCAACCUGGGGGGCAGUGGUGUGGAGAAGCCACAGGCCUGAGAUUUCAUCUGCUCUCCUUCCUGGAGCCCAGAGGAGGGCAGCAGAAGGGGGUGGUGUCUCCAACCCCCCAGCACUGAGGAAGAACGGGGCUCUUCUCAUUUCACCCCUCCCUUUCUCCCCUGCCCCCAGGACUGGGCCACUUCUGGGUGGGGCAGUGGGUCCCAGAUUGGCUCACACUGAGAAUGUAAGAACUACAAACAAAAUUUCUAUUAAAUUAAAUUUUGUGUCUCC  77 3′UTR-014CUCCCUCCAUCCCAACCUGGCUCCCUCCCACCCAACCAACUUUCCCCCCAACCCGG (Collal;AAACAGACAAGCAACCCAAACUGAACCCCCUCAAAAGCCAAAAAAUGGGAGACAAU collagen, TypeUUCACAUGGACUUUGGAAAAUAUUUUUUUCCUUUGCAUUCAUCUCUCAAACUUAGU I, alpha 1UUUUAUCUUUGACCAACCGAACAUGACCAAAAACCAAAAGUGCAUUCAACCUUACC UTR)AAAAAAAAAAAAAAAAAAAGAAUAAAUAAAUAACUUUUUAAAAAAGGAAGCUUGGUCCACUUGCUUGAAGACCCAUGCGGGGGUAAGUCCCUUUCUGCCCGUUGGGCUUAUGAAACCCCAAUGCUGCCCUUUCUGCUCCUUUCUCCACACCCCCCUUGGGGCCUCCCCUCCACUCCUUCCCAAAUCUGUCUCCCCAGAAGACACAGGAAACAAUGUAUUGUCUGCCCAGCAAUCAAAGGCAAUGCUCAAACACCCAAGUGGCCCCCACCCUCAGCCCGCUCCUGCCCGCCCAGCACCCCCAGGCCCUGGGGGACCUGGGGUUCUCAGACUGCCAAAGAAGCCUUGCCAUCUGGCGCUCCCAUGGCUCUUGCAACAUCUCCCCUUCGUUUUUGAGGGGGUCAUGCCGGGGGAGCCACCAGCCCCUCACUGGGUUCGGAGGAGAGUCAGGAAGGGCCACGACAAAGCAGAAACAUCGGAUUUGGGGAACGCGUGUCAAUCCCUUGUGCCGCAGGGCUGGGCGGGAGAGACUGUUCUGUUCCUUGUGUAACUGUGUUGCUGAAAGACUACCUCGUUCUUGUCUUGAUGUGUCACCGGGGCAACUGCCUGGGGGCGGGGAUGGGGGCAGGGUGGAAGCGGCUCCCCAUUUUAUACCAAAGGUGCUACAUCUAUGUGAUGGGUGGGGUGGGGAGGGAAUCACUGGUGCUAUAGAAAUUGAGAUGCCCCCCCAGGCCAGCAAAUGUUCCUUUUUGUUCAAAGUCUAUUUUUAUUCCUUGAUAUUUUUCUUUUUUUUUUUUUUUUUUUGUGGAUGGGGACUUGUGAAUUUUUCUAAAGGUGCUAUUUAACAUGGGAGGAGAGCGUGUGCGGCUCCAGCCCAGCCCGCUGCUCACUUUCCACCCUCUCUCCACCUGCCUCUGGCUUCUCAGGCCUCUGCUCUCCGACCUCUCUCCUCUGAAACCCUCCUCCACAGCUGCAGCCCAUCCUCCCGGCUCCCUCCUAGUCUGUCCUGCGUCCUCUGUCCCCGGGUUUCAGAGACAACUUCCCAAAGCACAAAGCAGUUUUUCCCCCUAGGGGUGGGAGGAAGCAAAAGACUCUGUACCUAUUUUGUAUGUGUAUAAUAAUUUGAGAUGUUUUUAAUUAUUUUGAUUGCUGGAAUAAAGCAUGUGGAAAUGACCCAAACAUAAUCCGCAGUGGCCUCCUAAUUUCCUUCUUUGGAGUUGGGGGAGGGGUAGACAUGGGGAAGGGGCUUUGGGGUGAUGGGCUUGCCUUCCAUUCCUGCCCUUUCCCUCCCCACUAUUCUCUUCUAGAUCCCUCCAUAACCCCACUCCCCUUUCUCUCACCCUUCUUAUACCGCAAACCUUUCUACUUCCUCUUUCAUUUUCUAUUCUUGCAAUUUCCUUGCACCUUUUCCAAAUCCUCUUCUCCCCUGCAAUACCAUACAGGCAAUCCACGUGCACAACACACACACACACUCUUCACAUCUGGGGUUGUCCAAACCUCAUACCCACUCCCCUUCAAGCCCAUCCACUCUCCACCCCCUGGAUGCCCUGCACUUGGUGGCGGUGGGAUGCUCAUGGAUACUGGGAGGGUGAGGGGAGUGGAACCCGUGAGGAGGACCUGGGGGCCUCUCCUUGAACUGACAUGAAGGGUCAUCUGGCCUCUGCUCCCUUCUCACCCACGCUGACCUCCUGCCGAAGGAGCAACGCAACAGGAGAGGGGUCUGCUGAGCCUGGCGAGGGUCUGGGAGGGACCAGGAGGAAGGCGUGCUCCCUGCUCGCUGUCCUGGCCCUGGGGGAGUGAGGGAGACAGACACCUGGGAGAGCUGUGGGGAAGGCACUCGCACCGUGCUCUUGGGAAGGAAGGAGACCUGGCCCUGCUCACCACGGACUGGGUGCCUCGACCUCCUGAAUCCCCAGAACACAACCCCCCUGGGCUGGGGUGGUCUGGGGAACCAUCGUGCCCCCGCCUCCCGCCUACUCCUUUUUAAGCUU  78 3′UTR-015UUGGCCAGGCCUGACCCUCUUGGACCUUUCUUCUUUGCCGACAACCACUGCCCAGC (Plod1;AGCCUCUGGGACCUCGGGGUCCCAGGGAACCCAGUCCAGCCUCCUGGCUGUUGACU procollagen-UCCCAUUGCUCUUGGAGCCACCAAUCAAAGAGAUUCAAAGAGAUUCCUGCAGGCCA lysine, 2-GAGGCGGAACACACCUUUAUGGCUGGGGCUCUCCGUGGUGUUCUGGACCCAGCCCC oxoglutarate 5-UGGAGACACCAUUCACUUUUACUGCUUUGUAGUGACUCGUGCUCUCCAACCUGUCU dioxygenase 1UCCUGAAAAACCAAGGCCCCCUUCCCCCACCUCUUCCAUGGGGUGAGACUUGAGCA UTR)GAACAGGGGCUUCCCCAAGUUGCCCAGAAAGACUGUCUGGGUGAGAAGCCAUGGCCAGAGCUUCUCCCAGGCACAGGUGUUGCACCAGGGACUUCUGCUUCAAGUUUUGGGGUAAAGACACCUGGAUCAGACUCCAAGGGCUGCCCUGAGUCUGGGACUUCUGCCUCCAUGGCUGGUCAUGAGAGCAAACCGUAGUCCCCUGGAGACAGCGACUCCAGAGAACCUCUUGGGAGACAGAAGAGGCAUCUGUGCACAGCUCGAUCUUCUACUUGCCUGUGGGGAGGGGAGUGACAGGUCCACACACCACACUGGGUCACCCUGUCCUGGAUGCCUCUGAAGAGAGGGACAGACCGUCAGAAACUGGAGAGUUUCUAUUAAAGGUCAUUUAAACCA  79 3′UTR-016UCCUCCGGGACCCCAGCCCUCAGGAUUCCUGAUGCUCCAAGGCGACUGAUGGGCGC (Nucb1;UGGAUGAAGUGGCACAGUCAGCUUCCCUGGGGGCUGGUGUCAUGUUGGGCUCCUGG nucleobindin 1GGCGGGGGCACGGCCUGGCAUUUCACGCAUUGCUGCCACCCCAGGUCCACCUGUCU UTR)CCACUUUCACAGCCUCCAAGUCUGUGGCUCUUCCCUUCUGUCCUCCGAGGGGCUUGCCUUCUCUCGUGUCCAGUGAGGUGCUCAGUGAUCGGCUUAACUUAGAGAAGCCCGCCCCCUCCCCUUCUCCGUCUGUCCCAAGAGGGUCUGCUCUGAGCCUGCGUUCCUAGGUGGCUCGGCCUCAGCUGCCUGGGUUGUGGCCGCCCUAGCAUCCUGUAUGCCCACAGCUACUGGAAUCCCCGCUGCUGCUCCGGGCCAAGCUUCUGGUUGAUUAAUGAGGGCAUGGGGUGGUCCCUCAAGACCUUCCCCUACCUUUUGUGGAACCAGUGAUGCCUCAAAGACAGUGUCCCCUCCACAGCUGGGUGCCAGGGGCAGGGGAUCCUCAGUAUAGCCGGUGAACCCUGAUACCAGGAGCCUGGGCCUCCCUGAACCCCUGGCUUCCAGCCAUCUCAUCGCCAGCCUCCUCCUGGACCUCUUGGCCCCCAGCCCCUUCCCCACACAGCCCCAGAAGGGUCCCAGAGCUGACCCCACUCCAGGACCUAGGCCCAGCCCCUCAGCCUCAUCUGGAGCCCCUGAAGACCAGUCCCACCCACCUUUCUGGCCUCAUCUGACACUGCUCCGCAUCCUGCUGUGUGUCCUGUUCCAUGUUCCGGUUCCAUCCAAAUACACUUUCUG GAACAAA  803′UTR-017 (α- GCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUglobin) CCCCUUCCUGCACCCGUACCCCCGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC  813′UTR-018 UGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCGUGGUCUUUGAAUAAAGUCUGAGU GGGCGGC  825′UTR-001 GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACC  83 5′UTR-002GGGAGAUCAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACC  84 5′UTR-003GGAAUAAAAGUCUCAACACAACAUAUACAAAACAAACGAAUCUCAAGCAAUCAAGCAUUCUACUUCUAUUGCAGCAAUUUAAAUCAUUUCUUUUAAAGCAAAAGCAAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUAGCAAC  85 5′UTR-004GGAGACAAGCUUGGCAUUCCGGUACUGUUGGUAAAGCCACC  86 5′UTR-005GGGAGAUCAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACC  87 5′UTR-006GGAAUAAAAGUCUCAACACAACAUAUACAAAACAAACGAAUCUCAAGCAAUCAAGCAUUCUACUUCUAUUGCAGCAAUUUAAAUCAUUUCUUUUAAAGCAAAAGCAAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUAGCAAC  88 5′UTR-007GGGAGACAAGCUUGGCAUUCCGGUACUGUUGGUAAAGCCACC  89 5′UTR-008GGGAAUUAACAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACC  90 5′UTR-009GGGAAAUUAGACAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACC  91 5′UTR-010GGGAAAUAAGAGAGUAAAGAACAGUAAGAAGAAAUAUAAGAGCCACC  92 5′UTR-011GGGAAAAAAGAGAGAAAAGAAGACUAAGAAGAAAUAUAAGAGCCACC  93 5′UTR-012GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAUAUAUAAGAGCCACC  94 5′UTR-013GGGAAAUAAGAGACAAAACAAGAGUAAGAAGAAAUAUAAGAGCCACC  95 5′UTR-014GGGAAAUUAGAGAGUAAAGAACAGUAAGUAGAAUUAAAAGAGCCACC  96 5′UTR-015GGGAAAUAAGAGAGAAUAGAAGAGUAAGAAGAAAUAUAAGAGCCACC  97 5′UTR-016GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAAUUAAGAGCCACC  98 hIL12AB_001TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGTCACCAGCAGCTGGT 3′UTR)CATTAGCTGGTTTAGCCTTGTGTTCCTGGCCTCCCCCCTTGTCGCTATTTGGGAGCTCAAGAAGGACGTGTACGTGGTGGAGTTGGATTGGTACCCAGACGCGCCCGGAGAGATGGTAGTTCTGACCTGTGATACCCCAGAGGAGGACGGCATCACCTGGACGCTGGACCAAAGCAGCGAGGTTTTGGGCTCAGGGAAAACGCTGACCATCCAGGTGAAGGAATTCGGCGACGCCGGGCAGTACACCTGCCATAAGGGAGGAGAGGTGCTGAGCCATTCCCTTCTTCTGCTGCACAAGAAAGAGGACGGCATCTGGTCTACCGACATCCTGAAAGACCAGAAGGAGCCCAAGAACAAAACCTTCCTGAGGTGCGAGGCCAAGAACTACTCCGGCAGGTTCACTTGTTGGTGGCTGACCACCATCAGTACAGACCTGACTTTTAGTGTAAAAAGCTCCAGAGGCTCGTCCGATCCCCAAGGGGTGACCTGCGGCGCAGCCACTCTGAGCGCTGAGCGCGTGCGCGGTGACAATAAAGAGTACGAGTACAGCGTTGAGTGTCAAGAAGATAGCGCTTGCCCTGCCGCCGAGGAGAGCCTGCCTATCGAGGTGATGGTTGACGCAGTGCACAAGCTTAAGTACGAGAATTACACCAGCTCATTCTTCATTAGAGATATAATCAAGCCTGACCCACCCAAGAACCTGCAGCTGAAGCCACTGAAAAACTCACGGCAGGTCGAAGTGAGCTGGGAGTACCCCGACACCTGGAGCACTCCTCATTCCTATTTCTCTCTTACATTCTGCGTCCAGGTGCAGGGCAAGAGCAAGCGGGAAAAGAAGGATCGAGTCTTCACCGACAAAACAAGCGCGACCGTGATTTGCAGGAAGAACGCCAGCATCTCCGTCAGAGCCCAGGATAGATACTATAGTAGCAGCTGGAGCGAGTGGGCAAGCGTGCCCTGTTCCGGCGGCGGGGGCGGGGGCAGCCGAAACTTGCCTGTCGCTACCCCGGACCCTGGAATGTTTCCGTGTCTGCACCACAGCCAGAACCTGCTGAGAGCCGTGTCGAATATGCTCCAGAAGGCCCGGCAGACCCTTGAGTTCTACCCCTGTACCAGCGAAGAGATCGATCATGAAGATATCACGAAAGATAAAACATCCACCGTCGAGGCTTGTCTCCCGCTGGAGCTGACCAAGAACGAGAGCTGTCTGAATAGCCGGGAGACGTCTTTCATCACGAATGGTAGCTGTCTGGCCAGCAGGAAAACTTCCTTCATGATGGCTCTCTGCCTGAGCTCTATCTATGAAGATCTGAAGATGTATCAGGTGGAGTTTAAAACAATGAACGCCAAACTCCTGATGGACCCAAAAAGGCAAATCTTTCTGGACCAGAATATGCTGGCCGTGATAGACGAGCTGATGCAGGCACTGAACTTCAACAGCGAGACGGTGCCACAGAAATCCAGCCTGGAGGAGCCTGACTTTTACAAAACTAAGATCAAGCTGTGTATCCTGCTGCACGCCTTTAGAATCCGTGCCGTGACTATCGACAGGGTGATGTCATACCTCAACGCTTCATGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC  99 hIL12AB_002TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5’UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAGCAGCTGGT 3’UTR)GATCAGCTGGTTCAGCCTGGTGTTCCTGGCCAGCCCCCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGTTGGATTGGTACCCCGACGCCCCCGGCGAGATGGTGGTGCTGACCTGCGACACCCCCGAGGAGGACGGCATCACCTGGACCCTGGACCAGAGCAGCGAGGTGCTGGGCAGCGGCAAGACCCTGACCATCCAGGTGAAGGAGTTCGGCGACGCCGGCCAGTACACCTGCCACAAGGGCGGCGAGGTGCTGAGCCACAGCCTGCTGCTGCTGCACAAGAAGGAGGACGGCATCTGGAGCACCGACATCCTGAAGGACCAGAAGGAGCCCAAGAACAAGACCTTCCTGAGATGCGAGGCCAAGAACTACAGCGGCAGATTCACCTGCTGGTGGCTGACCACCATCAGCACCGACCTGACCTTCAGCGTGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGTGACCTGCGGCGCCGCCACCCTGAGCGCCGAGAGAGTGAGAGGCGACAACAAGGAGTACGAGTACAGCGTGGAGTGCCAGGAAGATAGCGCCTGCCCCGCCGCCGAGGAGAGCCTGCCCATCGAGGTGATGGTGGACGCCGTGCACAAGCTGAAGTACGAGAACTACACCAGCAGCTTCTTCATCAGAGATATCATCAAGCCCGACCCCCCCAAGAACCTGCAGCTGAAGCCCCTGAAGAACAGCCGGCAGGTGGAGGTGAGCTGGGAGTACCCCGACACCTGGAGCACCCCCCACAGCTACTTCAGCCTGACCTTCTGCGTGCAGGTGCAGGGCAAGAGCAAGAGAGAGAAGAAAGATAGAGTGTTCACCGACAAGACCAGCGCCACCGTGATCTGCAGAAAGAACGCCAGCATCAGCGTGAGAGCCCAAGATAGATACTACAGCAGCAGCTGGAGCGAGTGGGCCAGCGTGCCCTGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCTGCCCGTGGCCACCCCCGACCCCGGCATGTTCCCCTGCCTGCACCACAGCCAGAACCTGCTGAGAGCCGTGAGCAACATGCTGCAGAAGGCCCGGCAGACCCTGGAGTTCTACCCCTGCACCAGCGAGGAGATCGACCACGAAGATATCACCAAAGATAAGACCAGCACCGTGGAGGCCTGCCTGCCCCTGGAGCTGACCAAGAACGAGAGCTGCCTGAACAGCAGAGAGACCAGCTTCATCACCAACGGCAGCTGCCTGGCCAGCAGAAAGACCAGCTTCATGATGGCCCTGTGCCTGAGCAGCATCTACGAGGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAACGCCAAGCTGCTGATGGACCCCAAGCGGCAGATCTTCCTGGACCAGAACATGCTGGCCGTGATCGACGAGCTGATGCAGGCCCTGAACTTCAACAGCGAGACCGTGCCCCAGAAGAGCAGCCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATCCTGCTGCACGCCTTCAGAATCAGAGCCGTGACCATCGACAGAGTGATGAGCTACCTGAACGCCAGCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 100 hIL12AB_003TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGTCACCAGCAGTTGGT 3′UTR)CATCTCTTGGTTTTCCCTGGTTTTTCTGGCATCTCCCCTCGTGGCCATCTGGGAACTGAAGAAAGACGTTTACGTTGTAGAATTGGATTGGTATCCGGACGCTCCTGGAGAAATGGTGGTCCTCACCTGTGACACCCCTGAAGAAGACGGAATCACCTGGACCTTGGACCAGAGCAGTGAGGTCTTAGGCTCTGGCAAAACCCTGACCATCCAAGTCAAAGAGTTTGGAGATGCTGGCCAGTACACCTGTCACAAAGGAGGCGAGGTTCTAAGCCATTCGCTCCTGCTGCTTCACAAAAAGGAAGATGGAATTTGGTCCACTGATATTTTAAAGGACCAGAAAGAACCCAAAAATAAGACCTTTCTAAGATGCGAGGCCAAGAATTATTCTGGACGTTTCACCTGCTGGTGGCTGACGACAATCAGTACTGATTTGACATTCAGTGTCAAAAGCAGCAGAGGCTCTTCTGACCCCCAAGGGGTGACGTGCGGAGCTGCTACACTCTCTGCAGAGAGAGTCAGAGGTGACAACAAGGAGTATGAGTACTCAGTGGAGTGCCAGGAAGATAGTGCCTGCCCAGCTGCTGAGGAGAGTCTGCCCATTGAGGTCATGGTGGATGCCGTTCACAAGCTCAAGTATGAAAACTACACCAGCAGCTTCTTCATCAGAGATATCATCAAACCTGACCCACCCAAGAACTTGCAGCTGAAGCCATTAAAGAATTCTCGGCAGGTGGAGGTCAGCTGGGAGTACCCTGACACCTGGAGTACTCCACATTCCTACTTCTCCCTGACATTCTGCGTTCAGGTCCAGGGCAAGAGCAAGAGAGAAAAGAAAGATAGAGTCTTCACAGATAAGACCTCAGCCACGGTCATCTGCCGCAAAAATGCCAGCATTAGCGTGCGGGCCCAGGACCGCTACTATAGCTCATCTTGGAGCGAATGGGCATCTGTGCCCTGCAGTGGCGGAGGGGGCGGAGGGAGCAGAAACCTCCCCGTGGCCACTCCAGACCCAGGAATGTTCCCATGCCTTCACCACTCCCAAAACCTGCTGAGGGCCGTCAGCAACATGCTCCAGAAGGCCCGGCAAACTTTAGAATTTTACCCTTGCACTTCTGAAGAGATTGATCATGAAGATATCACAAAAGATAAAACCAGCACAGTGGAGGCCTGTTTACCATTGGAATTAACCAAGAATGAGAGTTGCCTAAATTCCAGAGAGACCTCTTTCATAACTAATGGGAGTTGCCTGGCCTCCAGAAAGACCTCTTTTATGATGGCCCTGTGCCTTAGTAGTATTTATGAAGATTTGAAGATGTACCAGGTGGAGTTCAAGACCATGAATGCAAAGCTTCTGATGGATCCTAAGAGGCAGATCTTTTTAGATCAAAACATGCTGGCAGTTATTGATGAGCTGATGCAGGCCCTGAATTTCAACAGTGAGACGGTGCCACAAAAATCCTCCCTTGAAGAACCAGATTTCTACAAGACCAAGATCAAGCTCTGCATACTTCTTCATGCTTTCAGAATTCGGGCAGTGACTATTGATAGAGTGATGAGCTATCTGAATGCTTCCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 101 Human CD8AIYIWAPLAGTCGVLLLSLVITLYCY Transmembrane Domain 102 HumanVVVISAILALVVLTIISLIILIMLW PDGF-RB Transmembrane domain 103 Human CD80LLPSWAITLISVNGIFVICCL Transmembrane domain 104 hIL12AB_004TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGGGCTGCCACCAGCAGCT 3′UTR)GGTCATCAGCTGGTTCTCCCTGGTCTTCCTGGCCAGCCCCCTGGTGGCCATCTGGGAGCTGAAGAAAGACGTCTACGTAGTAGAGTTGGATTGGTACCCAGACGCACCTGGAGAAATGGTGGTTCTCACCTGTGACACGCCAGAAGAAGACGGTATCACCTGGACGCTGGACCAGAGCTCAGAAGTTCTTGGCAGTGGAAAAACGCTGACCATACAAGTAAAAGAATTTGGGGATGCTGGCCAGTACACCTGCCACAAAGGAGGAGAAGTTCTCAGCCACAGCCTGCTGCTGCTGCACAAGAAAGAAGATGGCATCTGGAGCACAGATATTTTAAAAGACCAGAAGGAGCCCAAGAACAAAACCTTCCTTCGATGTGAGGCCAAGAACTACAGTGGCCGCTTCACCTGCTGGTGGCTCACCACCATCAGCACAGACCTCACCTTCTCGGTGAAGAGCAGCCGTGGCAGCTCAGACCCCCAAGGAGTCACCTGTGGGGCGGCCACGCTGTCGGCAGAAAGAGTTCGAGGTGACAACAAGGAATATGAATACTCGGTGGAATGTCAAGAAGATTCGGCCTGCCCGGCGGCAGAAGAAAGTCTTCCCATAGAAGTCATGGTGGATGCTGTTCACAAATTAAAATATGAAAACTACACCAGCAGCTTCTTCATCAGAGATATCATCAAGCCAGACCCGCCCAAGAACCTGCAGCTGAAGCCCCTGAAGAACAGCCGGCAGGTGGAAGTTTCCTGGGAGTACCCAGATACGTGGAGCACGCCGCACAGCTACTTCAGCCTCACCTTGTGTGTACAAGTACAAGGCAAGAGCAAGAGAGAGAAGAAAGATCGTGTCTTCACAGATAAAACCTCGGCGACGGTCATCTGCAGGAAGAATGCCTCCATCTCGGTTCGAGCCCAGGACCGCTACTACAGCAGCAGCTGGAGTGAGTGGGCCTCGGTGCCCTGCAGTGGTGGCGGCGGCGGCGGCAGCAGAAACCTTCCTGTGGCCACGCCGGACCCTGGCATGTTCCCGTGCCTGCACCACAGCCAAAATTTACTTCGAGCTGTTTCTAACATGCTGCAGAAAGCACGGCAAACTTTAGAATTCTACCCCTGCACCTCAGAAGAAATAGACCATGAAGATATCACCAAAGATAAAACCAGCACTGTAGAGGCCTGCCTGCCCCTGGAGCTCACCAAGAATGAATCCTGCCTCAACAGCAGAGAGACCAGCTTCATCACCAATGGCAGCTGCCTGGCCAGCAGGAAAACCAGCTTCATGATGGCGCTCTGCCTGAGCAGCATCTATGAAGATTTGAAGATGTACCAAGTAGAATTTAAAACCATGAATGCCAAGCTGCTCATGGACCCCAAGCGGCAGATATTTTTGGATCAAAACATGCTGGCTGTCATTGATGAGCTCATGCAAGCATTAAACTTCAACTCAGAGACGGTGCCCCAGAAGAGCAGCCTGGAGGAGCCAGATTTCTACAAAACCAAGATCAAGCTCTGCATCTTATTACATGCCTTCCGCATCCGGGCGGTCACCATTGACCGTGTCATGTCCTACTTAAATGCCAGCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 105 hIL12AB_005TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAGCAGCTGGT 3′UTR)CATCAGCTGGTTCTCCCTGGTCTTCCTGGCCAGCCCCCTGGTGGCCATCTGGGAGCTGAAGAAAGACGTCTACGTAGTAGAGTTGGATTGGTACCCAGACGCACCTGGAGAAATGGTGGTTCTCACCTGTGACACGCCAGAAGAAGACGGTATCACCTGGACGCTGGACCAGAGCTCAGAAGTTCTTGGCAGTGGAAAAACGCTGACCATACAAGTAAAAGAATTTGGGGATGCTGGCCAGTACACCTGCCACAAAGGAGGAGAAGTTCTCAGCCACAGCCTGCTGCTGCTGCACAAGAAAGAAGATGGCATCTGGAGCACAGATATTTTAAAAGACCAGAAGGAGCCCAAGAACAAAACCTTCCTTCGATGTGAGGCCAAGAACTACAGTGGCCGCTTCACCTGCTGGTGGCTCACCACCATCAGCACAGACCTCACCTTCTCGGTGAAGAGCAGCCGTGGCAGCTCAGACCCCCAAGGAGTCACCTGTGGGGCGGCCACGCTGTCGGCAGAAAGAGTTCGAGGTGACAACAAGGAATATGAATACTCGGTGGAATGTCAAGAAGATTCGGCCTGCCCGGCGGCAGAAGAAAGTCTTCCCATAGAAGTCATGGTGGATGCTGTTCACAAATTAAAATATGAAAACTACACCAGCAGCTTCTTCATCAGAGATATCATCAAGCCAGACCCGCCCAAGAACCTGCAGCTGAAGCCCCTGAAGAACAGCCGGCAGGTGGAAGTTTCCTGGGAGTACCCAGATACGTGGAGCACGCCGCACAGCTACTTCAGCCTCACCTTCTGTGTACAAGTACAAGGCAAGAGCAAGAGAGAGAAGAAAGATCGTGTCTTCACAGATAAAACCTCGGCGACGGTCATCTGCAGGAAGAATGCCTCCATCTCGGTTCGAGCCCAGGACCGCTACTACAGCAGCAGCTGGAGTGAGTGGGCCTCGGTGCCCTGCAGTGGTGGCGGCGGCGGCGGCAGCAGAAACCTTCCTGTGGCCACGCCGGACCCTGGCATGTTCCCGTGCCTGCACCACAGCCAAAATTTACTTCGAGCTGTTTCTAACATGCTGCAGAAAGCACGGCAAACTTTAGAATTCTACCCCTGCACCTCAGAAGAAATAGACCATGAAGATATCACCAAAGATAAAACCAGCACTGTAGAGGCCTGCCTGCCCCTGGAGCTCACCAAGAATGAATCCTGCCTCAACAGCAGAGAGACCAGCTTCATCACCAATGGCAGCTGCCTGGCCAGCAGGAAAACCAGCTTCATGATGGCGCTCTGCCTGAGCAGCATCTATGAAGATTTGAAGATGTACCAAGTAGAATTTAAAACCATGAATGCCAAGCTGCTCATGGACCCCAAGCGGCAGATATTTTTGGATCAAAACATGCTGGCTGTCATTGATGAGCTCATGCAAGCATTAAACTTCAACTCAGAGACGGTGCCCCAGAAGAGCAGCCTGGAGGAGCCAGATTTCTACAAAACCAAGATCAAGCTCTGCATCTTATTACATGCCTTCCGCATCCGGGCGGTCACCATTGACCGTGTCATGTCCTACTTAAATGCCAGCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 106 hIL12AB_006TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAGCAGCTGGT 3′UTR)GATCAGCTGGTTCAGCCTGGTGTTCCTGGCCAGCCCCCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGTTGGATTGGTACCCCGACGCCCCCGGCGAGATGGTGGTGCTGACCTGTGACACCCCCGAGGAGGACGGCATCACCTGGACCCTGGACCAGAGCAGCGAGGTGCTGGGCAGCGGCAAGACCCTGACCATCCAGGTGAAGGAGTTCGGGGACGCCGGCCAGTACACCTGCCACAAGGGCGGCGAGGTGCTGAGCCACAGCCTGCTGCTGCTGCACAAGAAGGAGGACGGCATCTGGAGCACAGATATCCTGAAGGACCAGAAGGAGCCCAAGAACAAGACCTTCCTGAGATGCGAGGCCAAGAACTACAGCGGCAGATTCACCTGCTGGTGGCTGACCACCATCAGCACAGATTTGACCTTCAGCGTGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGTGACCTGCGGCGCCGCCACCCTGAGCGCCGAGAGAGTGAGAGGTGACAACAAGGAGTACGAGTACAGCGTGGAGTGCCAGGAAGATAGCGCCTGCCCCGCCGCCGAGGAGAGCCTGCCCATCGAGGTGATGGTGGACGCCGTGCACAAGCTGAAGTACGAGAACTACACCAGCAGCTTCTTCATCAGAGATATCATCAAGCCCGACCCGCCGAAGAACCTGCAGCTGAAGCCCCTGAAGAACAGCCGGCAGGTGGAGGTGAGCTGGGAGTACCCCGACACCTGGAGCACCCCCCACAGCTACTTCAGCCTGACCTTCTGCGTGCAGGTGCAGGGCAAGAGCAAGAGAGAGAAGAAAGATAGAGTGTTCACAGATAAGACCAGCGCCACCGTGATCTGCAGAAAGAACGCCAGCATCAGCGTGAGAGCCCAAGATAGATACTACAGCAGCAGCTGGAGCGAGTGGGCCAGCGTGCCCTGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCTGCCCGTGGCCACCCCCGACCCCGGCATGTTCCCCTGCCTGCACCACAGCCAGAACCTGCTGAGAGCCGTGAGCAACATGCTGCAGAAGGCCCGGCAGACCCTGGAGTTCTACCCCTGCACCAGCGAGGAGATCGACCACGAAGATATCACCAAAGATAAGACCAGCACCGTGGAGGCCTGCCTGCCCCTGGAGCTGACCAAGAATGAAAGCTGCCTGAACAGCAGAGAGACCAGCTTCATCACCAACGGCAGCTGCCTGGCCAGCAGAAAGACCAGCTTCATGATGGCCCTGTGCCTGAGCAGCATCTACGAGGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAACGCCAAGCTGCTGATGGACCCCAAGCGGCAGATCTTCCTGGACCAGAACATGCTGGCCGTGATCGACGAGCTGATGCAGGCCCTGAACTTCAACAGCGAGACCGTGCCCCAGAAGAGCAGCCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATCCTGCTGCACGCCTTCAGAATCAGAGCCGTGACCATCGACAGAGTGATGAGCTACCTGAACGCCAGCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 107 hIL12AB_007TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAGCAGCTTGT 3′UTR)CATCTCCTGGTTCTCTCTTGTCTTCCTTGCTTCTCCTCTTGTGGCCATCTGGGAGCTGAAGAAGGACGTTTACGTAGTGGAGTTGGATTGGTACCCTGACGCACCTGGAGAAATGGTGGTTCTCACCTGTGACACTCCTGAGGAGGACGGTATCACCTGGACGTTGGACCAGTCTTCTGAGGTTCTTGGCAGTGGAAAAACTCTTACTATTCAGGTGAAGGAGTTTGGAGATGCTGGCCAGTACACCTGCCACAAGGGTGGTGAAGTTCTCAGCCACAGTTTACTTCTTCTTCACAAGAAGGAGGATGGCATCTGGTCTACTGACATTTTAAAAGACCAGAAGGAGCCCAAGAACAAAACATTCCTTCGTTGTGAAGCCAAGAACTACAGTGGTCGTTTCACCTGCTGGTGGCTTACTACTATTTCTACTGACCTTACTTTCTCTGTGAAGTCTTCTCGTGGCTCTTCTGACCCTCAGGGTGTCACCTGTGGGGCTGCTACTCTTTCTGCTGAGCGTGTGCGTGGTGACAACAAGGAGTATGAATACTCGGTGGAGTGCCAGGAAGATTCTGCCTGCCCTGCTGCTGAGGAGTCTCTTCCTATTGAGGTGATGGTGGATGCTGTGCACAAGTTAAAATATGAAAACTACACTTCTTCTTTCTTCATTCGTGACATTATAAAACCTGACCCTCCCAAGAACCTTCAGTTAAAACCTTTAAAAAACTCTCGTCAGGTGGAGGTGTCCTGGGAGTACCCTGACACGTGGTCTACTCCTCACTCCTACTTCTCTCTTACTTTCTGTGTCCAGGTGCAGGGCAAGTCCAAGCGTGAGAAGAAGGACCGTGTCTTCACTGACAAAACATGTGCTACTGTCATCTGCAGGAAGAATGCATCCATCTCTGTGCGTGCTCAGGACCGTTACTACAGCTCTTCCTGGTCTGAGTGGGCTTCTGTGCCCTGCTCTGGCGGCGGCGGCGGCGGCAGCAGAAATCTTCCTGTGGCTACTCCTGACCCTGGCATGTTCCCCTGCCTTCACCACTCGCAGAACCTTCTTCGTGCTGTGAGCAACATGCTTCAGAAGGCTCGTCAAACTTTAGAATTCTACCCCTGCACTTCTGAGGAGATTGACCATGAAGATATCACCAAAGATAAAACATCTACTGTGGAGGCCTGCCTTCCTTTAGAGCTGACCAAGAATGAATCCTGCTTAAATTCTCGTGAGACGTCTTTCATCACCAATGGCAGCTGCCTTGCCTCGCGCAAAACATCTTTCATGATGGCTCTTTGCCTTTCTTCCATCTATGAAGATTTAAAAATGTACCAGGTGGAGTTCAAGACCATGAATGCAAAGCTTCTCATGGACCCCAAGCGTCAGATATTTTTGGACCAGAACATGCTTGCTGTCATTGATGAGCTCATGCAGGCTTTAAACTTCAACTCTGAGACGGTGCCTCAGAAGTCTTCTTTAGAAGAGCCTGACTTCTACAAGACCAAGATAAAACTTTGCATTCTTCTTCATGCTTTCCGCATCCGTGCTGTGACTATTGACCGTGTGATGTCCTACTTAAATGCTTCTTGATAATAGGCTGGAGCCTCGGTGGCCAAGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 108 hIL12AB_008TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGTCATCAACAACTCGT 3′UTR)GATTAGCTGGTTCAGTCTCGTGTTCCTGGCCTCTCCGCTGGTGGCCATCTGGGAGCTTAAGAAGGACGTGTACGTGGTGGAGCTCGATTGGTACCCCGACGCACCTGGCGAGATGGTGGTGCTAACCTGCGATACCCCCGAGGAGGACGGGATCACTTGGACCCTGGATCAGAGTAGCGAAGTCCTGGGCTCTGGCAAAACACTCACAATCCAGGTGAAGGAATTCGGAGACGCTGGTCAGTACACTTGCCACAAGGGGGGTGAAGTGCTGTCTCACAGCCTGCTGTTACTGCACAAGAAGGAGGATGGGATCTGGTCAACCGACATCCTGAAGGATCAGAAGGAGCCTAAGAACAAGACCTTTCTGAGGTGTGAAGCTAAGAACTATTCCGGAAGATTCACTTGCTGGTGGTTGACCACAATCAGCACTGACCTGACCTTTTCCGTGAAGTCCAGCAGAGGAAGCAGCGATCCTCAGGGCGTAACGTGCGGCGCGGCTACCCTGTCAGCTGAGCGGGTTAGAGGCGACAACAAAGAGTATGAGTACTCCGTGGAGTGTCAGGAAGATAGCGCCTGCCCCGCAGCCGAGGAGAGTCTGCCCATCGAGGTGATGGTGGACGCTGTCCATAAGTTAAAATACGAAAATTACACAAGTTCCTTTTTCATCCGCGATATTATCAAACCCGATCCCCCCAAGAACCTGCAGCTGAAGCCCCTGAAGAATAGCCGACAGGTGGAAGTCTCTTGGGAGTATCCTGACACCTGGTCCACGCCTCACAGCTACTTTAGTCTGACTTTCTGTGTCCAGGTCCAGGGCAAGAGCAAGAGAGAGAAAAAGGATAGAGTGTTTACTGACAAAACATCTGCTACAGTCATCTGCAGAAAGAACGCCAGTATCTCAGTGAGGGCGCAAGATAGATACTACAGTAGTAGCTGGAGCGAATGGGCTAGCGTGCCCTGTTCAGGGGGCGGCGGAGGGGGCTCCAGGAATCTGCCCGTGGCCACCCCCGACCCTGGGATGTTCCCTTGCCTCCATCACTCACAGAACCTGCTCAGAGCAGTGAGCAACATGCTCCAAAAGGCCCGCCAGACCCTGGAGTTTTACCCTTGTACTTCAGAAGAGATCGATCACGAAGATATAACAAAGGATAAAACCAGCACCGTGGAGGCCTGTCTGCCTCTGGAACTCACAAAGAATGAAAGCTGTCTGAATTCCAGGGAAACCTCCTTCATTACTAACGGAAGCTGTCTCGCATCTCGCAAAACATCATTCATGATGGCCCTCTGCCTGTCTTCTATCTATGAAGATCTCAAGATGTATCAGGTGGAGTTCAAAACAATGAACGCCAAGCTGCTGATGGACCCCAAGCGGCAGATCTTCCTGGACCAGAACATGCTGGCAGTGATCGATGAGCTGATGCAAGCCTTGAACTTCAACTCAGAGACGGTGCCGCAAAAGTCCTCGTTGGAGGAACCAGATTTTTACAAAACCAAAATCAAGCTGTGTATCCTTCTTCACGCCTTTCGGATCAGAGCCGTGACTATCGACCGGGTGATGTCATACCTGAATGCTTCCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 109 hIL12AB_009TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAGCAGCTGGT 3′UTR)CATCAGCTGGTTTAGCCTGGTCTTCCTGGCCAGCCCCCTGGTGGCCATCTGGGAGCTGAAGAAAGACGTCTACGTAGTAGAGTTGGATTGGTACCCAGACGCACCTGGAGAAATGGTGGTTCTCACCTGCGACACGCCAGAAGAAGACGGTATCACCTGGACGCTGGACCAGAGCAGCGAAGTACTGGGCAGTGGAAAAACGCTGACCATACAAGTAAAAGAATTTGGCGATGCTGGCCAGTACACCTGCCACAAAGGAGGAGAAGTACTGAGCCACAGCCTGCTGCTGCTGCACAAGAAAGAAGATGGCATCTGGAGCACCGACATTTTAAAAGACCAGAAGGAGCCCAAGAACAAAACCTTCCTTCGATGTGAGGCGAAGAACTACAGTGGCCGCTTCACCTGCTGGTGGCTCACCACCATCAGCACCGACCTCACCTTCTCGGTGAAGAGCAGCCGTGGTAGCTCAGACCCCCAAGGAGTCACCTGTGGGGCGGCCACGCTGTCGGCAGAAAGAGTTCGAGGCGACAACAAGGAATATGAATACTCGGTGGAATGTCAAGAAGATTCGGCCTGCCCGGCGGCAGAAGAAAGTCTGCCCATAGAAGTCATGGTGGATGCTGTTCACAAATTAAAATATGAAAACTACACCAGCAGCTTCTTCATCAGAGATATCATCAAGCCAGACCCCCCCAAGAACCTGCAGCTGAAGCCCCTGAAGAACAGCCGGCAGGTGGAAGTTTCCTGGGAGTACCCAGATACGTGGAGCACGCCGCACAGCTACTTCAGCCTCACCTTCTGTGTACAAGTACAAGGCAAGAGCAAGAGAGAGAAGAAAGATCGTGTCTTCACCGACAAAACCTCGGCGACGGTCATCTGCAGGAAGAATGCAAGCATCTCGGTTCGAGCCCAGGACCGCTACTACAGCAGCAGCTGGAGTGAGTGGGCCTCGGTGCCCTGCAGTGGTGGCGGCGGCGGCGGCAGCAGAAACCTTCCTGTGGCCACGCCGGACCCTGGCATGTTTCCGTGCCTGCACCACAGCCAAAATTTATTACGAGCTGTTAGCAACATGCTGCAGAAAGCACGGCAAACTTTAGAATTCTACCCCTGCACCTCAGAAGAAATAGACCATGAAGATATCACCAAAGATAAAACCAGCACTGTAGAGGCCTGCCTGCCCCTGGAGCTCACCAAGAACGAGAGCTGCCTCAATAGCAGAGAGACCAGCTTCATCACCAATGGCAGCTGCCTGGCCAGCAGGAAAACCAGCTTCATGATGGCGCTCTGCCTGAGCAGCATCTATGAAGATCTGAAGATGTACCAAGTAGAATTTAAAACCATGAATGCCAAGCTGCTCATGGACCCCAAGCGGCAGATATTCCTCGACCAAAACATGCTGGCTGTCATTGATGAGCTCATGCAAGCATTAAACTTCAACTCAGAGACGGTGCCCCAGAAGAGCAGCCTGGAGGAGCCAGATTTCTACAAAACCAAGATCAAGCTCTGCATCTTATTACATGCCTTCCGCATCCGGGCGGTCACCATTGACCGTGTCATGTCCTACTTAAATGCCAGCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 110 hIL12AB_010TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAGCAGCTTGT 3′UTR)CATCTCCTGGTTTTCTCTTGTCTTCCTCGCTTCTCCTCTTGTGGCCATCTGGGAGCTGAAGAAAGACGTCTACGTAGTAGAGTTGGATTGGTACCCGGACGCTCCTGGAGAAATGGTGGTTCTCACCTGCGACACTCCTGAAGAAGACGGTATCACCTGGACGCTGGACCAAAGCAGCGAAGTTTTAGGCTCTGGAAAAACGCTGACCATACAAGTAAAAGAATTTGGCGACGCTGGCCAGTACACGTGCCACAAAGGAGGAGAAGTTTTAAGCCACAGTTTACTTCTTCTTCACAAGAAAGAAGATGGCATCTGGAGTACAGATATTTTAAAAGACCAGAAGGAGCCTAAGAACAAAACCTTCCTCCGCTGTGAAGCTAAGAACTACAGTGGTCGTTTCACCTGCTGGTGGCTCACCACCATCTCCACTGACCTCACCTTCTCTGTAAAATCAAGCCGTGGTTCTTCTGACCCCCAAGGAGTCACCTGTGGGGCTGCCACGCTCAGCGCTGAAAGAGTTCGAGGCGACAACAAGGAATATGAATATTCTGTGGAATGTCAAGAAGATTCTGCCTGCCCGGCGGCAGAAGAAAGTCTTCCCATAGAAGTCATGGTGGACGCTGTTCACAAATTAAAATATGAAAACTACACCAGCAGCTTCTTCATTCGTGACATCATCAAACCAGACCCTCCTAAGAACCTTCAGTTAAAACCGCTGAAGAACAGCCGGCAGGTGGAAGTTTCCTGGGAGTACCCAGATACGTGGAGTACGCCGCACTCCTACTTCAGTTTAACCTTCTGTGTACAAGTACAAGGAAAATCAAAAAGAGAGAAGAAAGATCGTGTCTTCACTGACAAAACATCTGCCACGGTCATCTGCCGTAAGAACGCTTCCATCTCGGTTCGAGCCCAGGACCGCTACTACAGCAGCAGCTGGAGTGAGTGGGCATCTGTTCCCTGCAGTGGTGGCGGCGGCGGCGGCAGCCGCAACCTTCCTGTGGCCACGCCGGACCCTGGCATGTTCCCGTGCCTTCACCACTCGCAAAATCTTCTTCGTGCTGTTTCTAACATGCTGCAGAAGGCGCGGCAAACTTTAGAATTCTACCCGTGCACTTCTGAAGAAATAGACCATGAAGATATCACCAAAGATAAAACCAGCACGGTGGAGGCCTGCCTTCCTTTAGAACTTACTAAGAACGAAAGTTGCCTTAACAGCCGTGAGACCAGCTTCATCACCAATGGCAGCTGCCTTGCTAGCAGGAAGACCAGCTTCATGATGGCGCTGTGCCTTTCTTCCATCTATGAAGATCTTAAGATGTACCAAGTAGAATTTAAAACCATGAATGCCAAATTATTAATGGACCCCAAGCGGCAGATATTCCTCGACCAAAACATGCTGGCTGTCATTGATGAGCTCATGCAAGCATTAAACTTCAACTCAGAAACTGTTCCCCAGAAGTCATCTTTAGAAGAACCAGATTTCTACAAAACAAAAATAAAACTCTGCATTCTTCTTCATGCCTTCCGCATCCGTGCTGTCACCATTGACCGTGTCATGTCCTACTTAAATGCTTCTTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 111 hIL12AB_011TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAGCAGCTGGT 3′UTR)GATCAGCTGGTTCAGCCTGGTGTTCCTGGCCAGCCCCCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGTTGGATTGGTACCCGGACGCGCCGGGGGAGATGGTGGTGCTGACGTGCGACACGCCGGAGGAGGACGGGATCACGTGGACGCTGGACCAGAGCAGCGAGGTGCTGGGGAGCGGGAAGACGCTGACGATCCAGGTGAAGGAGTTCGGGGACGCGGGGCAGTACACGTGCCACAAGGGGGGGGAGGTGCTGAGCCACAGCCTGCTGCTGCTGCACAAGAAGGAGGACGGGATCTGGAGCACAGATATCCTGAAGGACCAGAAGGAGCCGAAGAACAAGACGTTCCTGAGGTGCGAGGCGAAGAACTACAGCGGGAGGTTCACGTGCTGGTGGCTGACGACGATCAGCACGGACCTGACGTTCAGCGTGAAGAGCAGCAGGGGGAGCAGCGACCCGCAGGGGGTGACGTGCGGGGCGGCGACGCTGAGCGCGGAGAGGGTGAGGGGTGACAACAAGGAGTACGAGTACAGCGTGGAGTGCCAGGAAGATAGCGCGTGCCCGGCGGCGGAGGAGAGCCTGCCGATCGAGGTGATGGTGGACGCGGTGCACAAGCTGAAGTACGAGAACTACACGAGCAGCTTCTTCATCAGAGATATCATCAAGCCGGACCCGCCGAAGAACCTGCAGCTGAAGCCGCTGAAGAACAGCAGGCAGGTGGAGGTGAGCTGGGAGTACCCAGATACGTGGAGCACGCCGCACAGCTACTTCAGCCTGACGTTCTGCGTGCAGGTGCAGGGGAAGAGCAAGAGGGAGAAGAAAGATAGGGTGTTCACAGATAAGACGAGCGCGACGGTGATCTGCAGGAAGAACGCGAGCATCAGCGTGAGGGCGCAAGATAGGTACTACAGCAGCAGCTGGAGCGAGTGGGCGAGCGTGCCGTGCAGCGGGGGGGGGGGGGGGGGGAGCAGGAACCTGCCGGTGGCGACGCCGGACCCGGGGATGTTCCCGTGCCTGCACCACAGCCAGAACCTGCTGAGGGCGGTGAGCAACATGCTGCAGAAGGCGAGGCAGACGCTGGAGTTCTACCCGTGCACGAGCGAGGAGATCGACCACGAAGATATCACGAAAGATAAGACGAGCACGGTGGAGGCGTGCCTGCCGCTGGAGCTGACGAAGAACGAGAGCTGCCTGAACAGCAGGGAGACGAGCTTCATCACGAACGGGAGCTGCCTGGCGAGCAGGAAGACGAGCTTCATGATGGCGCTGTGCCTGAGCAGCATCTACGAGGACCTGAAGATGTACCAGGTGGAGTTCAAGACGATGAACGCGAAGCTGCTGATGGACCCGAAGAGGCAGATCTTCCTGGACCAGAACATGCTGGCGGTGATCGACGAGCTGATGCAGGCGCTGAACTTCAACAGCGAGACGGTGCCGCAGAAGAGCAGCCTGGAGGAGCCAGATTTCTACAAGACGAAGATCAAGCTGTGCATCCTGCTGCACGCGTTCAGGATCAGGGCGGTGACGATCGACAGGGTGATGAGCTACCTGAACGCGAGCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 112 hIL12AB_012TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCATCAGCAGCTGGT 3′UTR)GATCAGCTGGTTCAGCCTCGTGTTTCTGGCCAGCCCCCTGGTGGCCATTTGGGAACTCAAGAAGGACGTGTACGTTGTGGAACTCGACTGGTACCCTGACGCCCCAGGCGAAATGGTGGTCTTAACCTGCGACACCCCTGAGGAGGACGGAATCACCTGGACCTTGGACCAGAGCTCCGAGGTCCTCGGCAGTGGCAAGACCCTGACCATACAGGTGAAAGAATTTGGAGACGCAGGGCAATACACATGTCACAAGGGCGGGGAGGTTCTTTCTCACTCCCTTCTGCTTCTACATAAAAAGGAAGACGGAATTTGGTCTACCGACATCCTCAAGGACCAAAAGGAGCCTAAGAATAAAACCTTCTTACGCTGTGAAGCTAAAAACTACAGCGGCAGATTCACTTGCTGGTGGCTCACCACCATTTCTACCGACCTGACCTTCTCGGTGAAGTCTTCAAGGGGCTCTAGTGATCCACAGGGAGTGACATGCGGGGCCGCCACACTGAGCGCTGAACGGGTGAGGGGCGATAACAAGGAGTATGAATACTCTGTCGAGTGTCAGGAGGATTCAGCTTGTCCCGCAGCTGAAGAGTCACTCCCCATAGAGGTTATGGTCGATGCTGTGCATAAACTGAAGTACGAAAACTACACCAGCAGCTTCTTCATTAGAGATATTATAAAACCTGACCCCCCCAAGAACCTGCAACTTAAACCCCTGAAAAACTCTCGGCAGGTCGAAGTTAGCTGGGAGTACCCTGATACTTGGTCCACCCCCCACTCGTACTTCTCACTGACTTTCTGTGTGCAGGTGCAGGGCAAGAGCAAGAGAGAGAAAAAAGATCGTGTATTCACAGATAAGACCTCTGCCACCGTGATCTGCAGAAAAAACGCTTCCATCAGTGTCAGAGCCCAAGACCGGTACTATAGTAGTAGCTGGAGCGAGTGGGCAAGTGTCCCCTGCTCTGGCGGCGGAGGGGGCGGCTCTCGAAACCTCCCCGTCGCTACCCCTGATCCAGGAATGTTCCCTTGCCTGCATCACTCACAGAATCTGCTGAGAGCGGTCAGCAACATGCTGCAGAAAGCTAGGCAAACACTGGAGTTTTATCCTTGTACCTCAGAGGAGATCGACCACGAGGATATTACCAAAGATAAGACCAGCACGGTGGAGGCCTGCTTGCCCCTGGAACTGACAAAGAATGAATCCTGCCTTAATAGCCGTGAGACCTCTTTTATAACAAACGGATCCTGCCTGGCCAGCAGGAAGACCTCCTTCATGATGGCCCTCTGCCTGTCCTCAATCTACGAAGACCTGAAGATGTACCAGGTGGAATTTAAAACTATGAACGCCAAGCTGTTGATGGACCCCAAGCGGCAGATCTTTCTGGATCAAAATATGCTGGCTGTGATCGACGAACTGATGCAGGCCCTCAACTTTAACAGCGAGACCGTGCCACAAAAGAGCAGTCTTGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATCCTCCTTCATGCCTTCAGGATAAGAGCTGTCACCATCGACAGAGTCATGAGTTACCTGAATGCATCCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 113 hIL12AB_013TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAGCAGCTGGT 3′UTR)CATCTCCTGGTTCAGTCTTGTCTTCCTGGCCTCGCCGCTGGTGGCCATCTGGGAGCTGAAGAAAGACGTTTACGTAGTAGAGTTGGATTGGTACCCAGACGCACCTGGAGAAATGGTGGTCCTCACCTGTGACACGCCAGAAGAAGACGGTATCACCTGGACGCTGGACCAGAGCAGTGAAGTTCTTGGAAGTGGAAAAACGCTGACCATACAAGTAAAAGAATTTGGAGATGCTGGCCAGTACACCTGCCACAAAGGAGGAGAAGTTCTCAGCCACAGTTTATTATTACTTCACAAGAAAGAAGATGGCATCTGGTCCACAGATATTTTAAAAGACCAGAAGGAGCCCAAAAATAAAACATTTCTTCGATGTGAGGCCAAGAACTACAGTGGTCGTTTCACCTGCTGGTGGCTGACCACCATCTCCACAGACCTCACCTTCAGTGTAAAAAGCAGCCGTGGTTCTTCTGACCCCCAAGGAGTCACCTGTGGGGCTGCCACGCTCTCTGCAGAAAGAGTTCGAGGTGACAACAAAGAATATGAGTACTCGGTGGAATGTCAAGAAGATTCGGCCTGCCCAGCTGCTGAGGAGAGTCTTCCCATAGAAGTCATGGTGGATGCTGTTCACAAATTAAAATATGAAAACTACACCAGCAGCTTCTTCATCAGAGATATCATCAAACCTGACCCGCCCAAGAACTTACAGCTGAAGCCGCTGAAAAACAGCCGGCAGGTAGAAGTTTCCTGGGAGTACCCAGATACCTGGTCCACGCCGCACTCCTACTTCTCCCTCACCTTCTGTGTACAAGTACAAGGCAAGAGCAAGAGAGAGAAGAAAGATCGTGTCTTCACAGATAAAACATCAGCCACGGTCATCTGCAGGAAAAATGCCAGCATCTCGGTGCGGGCCCAGGACCGCTACTACAGCAGCAGCTGGAGTGAGTGGGCATCTGTGCCCTGCAGTGGTGGTGGGGGTGGTGGCAGCAGAAACCTTCCTGTGGCCACTCCAGACCCTGGCATGTTCCCGTGCCTTCACCACTCCCAAAATTTACTTCGAGCTGTTTCTAACATGCTGCAGAAAGCACGGCAAACTTTAGAATTCTACCCGTGCACTTCTGAAGAAATTGACCATGAAGATATCACAAAAGATAAAACCAGCACAGTGGAGGCCTGTCTTCCTTTAGAGCTGACCAAAAATGAATCCTGCCTCAACAGCAGAGAGACCAGCTTCATCACCAATGGCAGCTGCCTGGCCTCCAGGAAAACCAGCTTCATGATGGCGCTCTGCCTCAGCTCCATCTATGAAGATTTGAAGATGTACCAAGTAGAATTTAAAACCATGAATGCCAAATTATTAATGGACCCCAAGAGGCAGATATTTTTAGATCAAAACATGCTGGCAGTTATTGATGAGCTCATGCAAGCATTAAACTTCAACAGTGAGACGGTACCTCAAAAAAGCAGCCTTGAAGAGCCAGATTTCTACAAAACCAAGATCAAACTCTGCATTTTACTTCATGCCTTCCGCATCCGGGCGGTCACCATTGACCGTGTCATGTCCTACTTAAATGCCTCGTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 114 hIL12AB_014TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAGCAGCTTGT 3′UTR)GATTTCTTGGTTCTCTCTTGTGTTCCTTGCTTCTCCTCTTGTGGCTATTTGGGAGTTAAAAAAGGACGTGTACGTGGTGGAGCTTGACTGGTACCCTGACGCACCTGGCGAGATGGTGGTGCTTACTTGTGACACTCCTGAGGAGGACGGCATTACTTGGACGCTTGACCAGTCTTCTGAGGTGCTTGGCTCTGGCAAAACACTTACTATTCAGGTGAAGGAGTTCGGGGATGCTGGCCAGTACACTTGCCACAAGGGCGGCGAGGTGCTTTCTCACTCTCTTCTTCTTCTTCACAAGAAGGAGGACGGCATTTGGTCTACTGACATTTTAAAAGACCAGAAGGAGCCCAAGAACAAAACATTCCTTCGTTGCGAGGCCAAGAACTACTCTGGCCGTTTCACTTGCTGGTGGCTTACTACTATTTCTACTGACCTTACTTTCTCTGTGAAGTCTTCTCGTGGCTCTTCTGACCCTCAGGGCGTGACTTGTGGGGCTGCTACTCTTTCTGCTGAGCGTGTGCGTGGTGACAACAAGGAGTACGAGTACTCTGTGGAGTGCCAGGAAGATTCTGCTTGCCCTGCTGCTGAGGAGTCTCTTCCTATTGAGGTGATGGTGGATGCTGTGCACAAGTTAAAATACGAGAACTACACTTCTTCTTTCTTCATTCGTGACATTATTAAGCCTGACCCTCCCAAGAACCTTCAGTTAAAACCTTTAAAAAACTCTCGTCAGGTGGAGGTGTCTTGGGAGTACCCTGACACTTGGTCTACTCCTCACTCTTACTTCTCTCTTACTTTCTGCGTGCAGGTGCAGGGCAAGTCTAAGCGTGAGAAGAAGGACCGTGTGTTCACTGACAAAACATCTGCTACTGTGATTTGCAGGAAGAATGCATCTATTTCTGTGCGTGCTCAGGACCGTTACTACTCTTCTTCTTGGTCTGAGTGGGCTTCTGTGCCTTGCTCTGGCGGCGGCGGCGGCGGCTCCAGAAATCTTCCTGTGGCTACTCCTGACCCTGGCATGTTCCCTTGCCTTCACCACTCTCAGAACCTTCTTCGTGCTGTGAGCAACATGCTTCAGAAGGCTCGTCAAACTCTTGAGTTCTACCCTTGCACTTCTGAGGAGATTGACCACGAAGATATCACCAAAGATAAAACATCTACTGTGGAGGCTTGCCTTCCTCTTGAGCTTACCAAGAATGAATCTTGCTTAAATTCTCGTGAGACGTCTTTCATCACCAACGGCTCTTGCCTTGCCTCGCGCAAAACATCTTTCATGATGGCTCTTTGCCTTTCTTCTATTTACGAAGATTTAAAAATGTACCAGGTGGAGTTCAAAACAATGAATGCAAAGCTTCTTATGGACCCCAAGCGTCAGATTTTCCTTGACCAGAACATGCTTGCTGTGATTGACGAGCTTATGCAGGCTTTAAATTTCAACTCTGAGACGGTGCCTCAGAAGTCTTCTCTTGAGGAGCCTGACTTCTACAAGACCAAGATTAAGCTTTGCATTCTTCTTCATGCTTTCCGTATTCGTGCTGTGACTATTGACCGTGTGATGTCTTACTTAAATGCTTCTTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 115 hIL12AB_015TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGTCACCAGCAGCTGGT 3′UTR)GATCAGCTGGTTTAGCCTGGTGTTTCTGGCCAGCCCCCTGGTGGCCATCTGGGAACTGAAGAAAGACGTGTACGTGGTAGAACTGGATTGGTATCCGGACGCTCCCGGCGAAATGGTGGTGCTGACCTGTGACACCCCCGAAGAAGACGGAATCACCTGGACCCTGGACCAGAGCAGCGAGGTGCTGGGCAGCGGCAAAACCCTGACCATCCAAGTGAAAGAGTTTGGCGATGCCGGCCAGTACACCTGTCACAAAGGCGGCGAGGTGCTAAGCCATTCGCTGCTGCTGCTGCACAAAAAGGAAGATGGCATCTGGAGCACCGATATCCTGAAGGACCAGAAAGAACCCAAAAATAAGACCTTTCTAAGATGCGAGGCCAAGAATTATAGCGGCCGTTTCACCTGCTGGTGGCTGACGACCATCAGCACCGATCTGACCTTCAGCGTGAAAAGCAGCAGAGGCAGCAGCGACCCCCAAGGCGTGACGTGCGGCGCCGCCACCCTGAGCGCCGAGAGAGTGAGAGGCGACAACAAGGAGTATGAGTACAGCGTGGAGTGCCAGGAAGATAGCGCCTGCCCCGCCGCCGAGGAGAGCCTGCCCATCGAGGTGATGGTGGATGCCGTGCACAAGCTGAAGTATGAAAACTACACCAGCAGCTTCTTCATCAGAGATATCATCAAACCCGACCCCCCCAAGAACCTGCAGCTGAAGCCCCTGAAGAATAGCCGGCAGGTGGAGGTGAGCTGGGAGTACCCCGACACCTGGAGCACCCCCCATAGCTACTTCAGCCTGACCTTCTGCGTGCAGGTGCAGGGCAAGAGCAAGAGAGAAAAGAAAGATAGAGTGTTCACAGATAAGACCAGCGCCACGGTGATCTGCAGAAAAAATGCCAGCATCAGCGTGAGAGCCCAAGATAGATACTATAGCAGCAGCTGGAGCGAATGGGCCAGCGTGCCCTGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCTGCCCGTGGCCACCCCCGACCCCGGCATGTTCCCCTGCCTGCACCACAGCCAAAACCTGCTGAGAGCCGTGAGCAACATGCTGCAGAAGGCCCGGCAGACCCTGGAATTTTACCCCTGCACCAGCGAAGAGATCGATCATGAAGATATCACCAAAGATAAAACCAGCACCGTGGAGGCCTGTCTGCCCCTGGAACTGACCAAGAATGAGAGCTGCCTAAATAGCAGAGAGACCAGCTTCATAACCAATGGCAGCTGCCTGGCCAGCAGAAAGACCAGCTTTATGATGGCCCTGTGCCTGAGCAGCATCTATGAAGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAATGCCAAGCTGCTGATGGATCCCAAGCGGCAGATCTTTCTGGATCAAAACATGCTGGCCGTGATCGATGAGCTGATGCAGGCCCTGAATTTCAACAGCGAGACCGTGCCCCAAAAAAGCAGCCTGGAAGAACCGGATTTTTATAAAACCAAAATCAAGCTGTGCATACTGCTGCATGCCTTCAGAATCAGAGCCGTGACCATCGATAGAGTGATGAGCTATCTGAATGCCAGCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 116 hIL12AB_016TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAGCAGCTGGT 3′UTR)CATCAGCTGGTTCAGCCTGGTCTTCCTGGCCAGCCCCCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTATACGTAGTGGAGTTGGATTGGTACCCAGACGCTCCTGGGGAGATGGTGGTGCTGACCTGTGACACCCCAGAAGAGGACGGTATCACCTGGACCCTGGACCAGAGCTCAGAAGTGCTGGGCAGTGGAAAAACCCTGACCATCCAGGTGAAGGAGTTTGGAGATGCTGGCCAGTACACCTGCCACAAGGGTGGTGAAGTGCTGAGCCACAGCCTGCTGCTGCTGCACAAGAAGGAGGATGGCATCTGGAGCACAGATATCCTGAAGGACCAGAAGGAGCCCAAGAACAAGACCTTCCTTCGCTGTGAAGCCAAGAACTACAGTGGCCGCTTCACCTGCTGGTGGCTGACCACCATCAGCACAGACCTCACCTTCTCGGTGAAGAGCAGCAGAGGCAGCTCAGACCCCCAGGGTGTCACCTGTGGGGCGGCCACGCTGTCGGCGGAGAGAGTTCGAGGTGACAACAAGGAGTATGAATACTCGGTGGAGTGCCAGGAAGATTCGGCGTGCCCGGCGGCAGAAGAGAGCCTGCCCATAGAAGTGATGGTGGATGCTGTGCACAAGCTGAAGTATGAAAACTACACCAGCAGCTTCTTCATCAGAGATATCATCAAGCCAGACCCGCCCAAGAACCTGCAGCTGAAGCCCCTGAAGAACAGCCGGCAGGTGGAGGTTTCCTGGGAGTACCCAGATACGTGGAGCACCCCCCACAGCTACTTCAGCCTGACCTTCTGTGTCCAGGTGCAGGGCAAGAGCAAGAGAGAGAAGAAAGATAGAGTCTTCACAGATAAGACCTCGGCCACGGTCATCTGCAGAAAGAATGCCTCCATCTCGGTTCGAGCCCAAGATAGATACTACAGCAGCAGCTGGTCAGAATGGGCCTCGGTGCCCTGCAGTGGTGGCGGCGGCGGCGGCAGCAGAAACCTGCCTGTTGCCACCCCAGACCCTGGGATGTTCCCCTGCCTGCACCACAGCCAGAACTTATTACGAGCTGTTTCTAACATGCTGCAGAAGGCCCGGCAGACCCTGGAGTTCTACCCCTGCACCTCAGAAGAGATTGACCATGAAGATATCACCAAAGATAAGACCAGCACTGTAGAGGCCTGCCTGCCCCTGGAGCTGACCAAGAATGAAAGCTGCCTGAACAGCAGAGAGACCAGCTTCATCACCAATGGAAGCTGCCTGGCCAGCAGAAAGACCAGCTTCATGATGGCCCTGTGCCTGAGCAGCATCTATGAAGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAATGCAAAGCTGCTGATGGACCCCAAGCGGCAGATATTTTTGGACCAGAACATGCTGGCTGTCATTGATGAGCTGATGCAGGCCCTGAACTTCAACTCAGAAACTGTACCCCAGAAGAGCAGCCTGGAGGAGCCAGATTTCTACAAGACCAAGATCAAGCTGTGCATCCTGCTTCATGCTTTCAGAATCAGAGCTGTCACCATTGACCGCGTGATGAGCTACTTAAATGCCTCGTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 117 hIL12AB_017TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAGCAGCTGGT 3′UTR)AATCAGCTGGTTTTCCCTCGTCTTTCTGGCATCACCCCTGGTGGCTATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGCTGGATTGGTACCCTGACGCCCCGGGGGAAATGGTGGTGTTAACCTGCGACACGCCTGAGGAGGACGGCATCACCTGGACGCTGGACCAGAGCAGCGAGGTGCTTGGGTCTGGTAAAACTCTGACTATTCAGGTGAAAGAGTTCGGGGATGCCGGCCAATATACTTGCCACAAGGGTGGCGAGGTGCTTTCTCATTCTCTGCTCCTGCTGCACAAGAAAGAAGATGGCATTTGGTCTACTGATATTCTGAAAGACCAGAAGGAGCCCAAGAACAAGACCTTTCTGAGATGCGAGGCTAAAAACTACAGCGGAAGATTTACCTGCTGGTGGCTGACCACAATCTCAACCGACCTGACATTTTCAGTGAAGTCCAGCAGAGGGAGCTCCGACCCTCAGGGCGTGACCTGCGGAGCCGCCACTCTGTCCGCAGAAAGAGTGAGAGGTGATAATAAGGAGTACGAGTATTCAGTCGAGTGCCAAGAAGATTCTGCCTGCCCAGCCGCCGAGGAGAGCCTGCCAATCGAGGTGATGGTAGATGCGGTACACAAGCTGAAGTATGAGAACTACACATCCTCCTTCTTCATAAGAGATATTATCAAGCCTGACCCACCTAAAAATCTGCAACTCAAGCCTTTGAAAAATTCACGGCAGGTGGAGGTGAGCTGGGAGTACCCTGATACTTGGAGCACCCCCCATAGCTACTTTTCGCTGACATTCTGCGTCCAGGTGCAGGGCAAGTCAAAGAGAGAGAAGAAGGATCGCGTGTTCACTGATAAAACAAGCGCCACAGTGATCTGCAGAAAAAACGCTAGCATTAGCGTCAGAGCACAGGACCGGTATTACTCCAGCTCCTGGAGCGAATGGGCATCTGTGCCCTGCAGCGGTGGGGGCGGAGGCGGATCCAGAAACCTCCCCGTTGCCACACCTGATCCTGGAATGTTCCCCTGTCTGCACCACAGCCAGAACCTGCTGAGAGCAGTGTCTAACATGCTCCAGAAGGCCAGGCAGACCCTGGAGTTTTACCCCTGCACCAGCGAGGAAATCGATCACGAAGATATCACCAAAGATAAAACCTCCACCGTGGAGGCCTGCCTGCCCCTGGAACTGACCAAAAACGAGAGCTGCCTGAATAGCAGGGAGACCTCCTTCATCACCAACGGCTCATGCCTTGCCAGCCGGAAAACTAGCTTCATGATGGCCCTGTGCCTGTCTTCGATCTATGAGGACCTGAAAATGTACCAGGTCGAATTTAAGACGATGAACGCAAAGCTGCTGATGGACCCCAAGCGGCAGATCTTTCTGGACCAGAACATGCTGGCAGTCATAGATGAGTTGATGCAGGCATTAAACTTCAACAGCGAGACCGTGCCTCAGAAGTCCAGCCTCGAGGAGCCAGATTTTTATAAGACCAAGATCAAACTATGCATCCTGCTGCATGCTTTCAGGATTAGAGCCGTCACCATCGATCGAGTCATGTCTTACCTGAATGCTAGCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 118 hIL12AB_018TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGTCACCAACAGTTAGT 3′UTR)AATCTCCTGGTTTTCTCTGGTGTTTCTGGCCAGCCCCCTCGTGGCCATCTGGGAGCTTAAAAAGGACGTTTACGTGGTGGAGTTGGATTGGTATCCCGACGCTCCAGGCGAAATGGTCGTGCTGACCTGCGATACCCCTGAAGAAGACGGTATCACCTGGACGCTGGACCAGTCTTCCGAGGTGCTTGGATCTGGCAAAACACTGACAATACAAGTTAAGGAGTTCGGGGACGCAGGGCAGTACACCTGCCACAAAGGCGGCGAGGTCCTGAGTCACTCCCTGTTACTGCTCCACAAGAAAGAGGACGGCATTTGGTCCACCGACATTCTGAAGGACCAGAAGGAGCCTAAGAATAAAACTTTCCTGAGATGCGAGGCAAAAAACTATAGCGGCCGCTTTACTTGCTGGTGGCTTACAACAATCTCTACCGATTTAACTTTCTCCGTGAAGTCTAGCAGAGGATCCTCTGACCCGCAAGGAGTGACTTGCGGAGCCGCCACCTTGAGCGCCGAAAGAGTCCGTGGCGATAACAAAGAATACGAGTACTCCGTGGAGTGCCAGGAAGATTCCGCCTGCCCAGCTGCCGAGGAGTCCCTGCCCATTGAAGTGATGGTGGATGCCGTCCACAAGCTGAAGTACGAAAACTATACCAGCAGCTTCTTCATCCGGGATATCATTAAGCCCGACCCTCCTAAAAACCTGCAACTTAAGCCCCTAAAGAATAGTCGGCAGGTTGAGGTCAGCTGGGAATATCCTGACACATGGAGCACCCCCCACTCTTATTTCTCCCTGACCTTCTGCGTGCAGGTGCAGGGCAAGAGTAAACGGGAGAAAAAAGATAGGGTCTTTACCGATAAAACCAGCGCTACGGTTATCTGTCGGAAGAACGCTTCCATCTCCGTCCGCGCTCAGGATCGTTACTACTCGTCCTCATGGAGCGAGTGGGCCAGCGTGCCCTGCAGCGGCGGCGGTGGAGGCGGATCCAGAAATCTGCCTGTTGCCACACCAGACCCTGGCATGTTCCCCTGTCTGCATCATAGCCAGAACCTGCTCAGAGCCGTGAGCAACATGCTCCAGAAGGCCAGGCAAACTTTGGAGTTCTACCCGTGTACATCTGAGGAAATCGATCACGAAGATATAACCAAAGATAAAACCTCTACAGTAGAGGCTTGTTTGCCCCTGGAGTTGACCAAAAACGAGAGTTGCCTGAACAGTCGCGAGACGAGCTTCATTACTAACGGCAGCTGTCTCGCCTCCAGAAAAACATCCTTCATGATGGCCCTGTGTCTTTCCAGCATATACGAAGACCTGAAAATGTACCAGGTCGAGTTCAAAACAATGAACGCCAAGCTGCTTATGGACCCCAAGCGGCAGATCTTCCTCGACCAAAACATGCTCGCTGTGATCGATGAGCTGATGCAGGCTCTCAACTTCAATTCCGAAACAGTGCCACAGAAGTCCAGTCTGGAAGAACCCGACTTCTACAAGACCAAGATTAAGCTGTGTATTTTGCTGCATGCGTTTAGAATCAGAGCCGTGACCATTGATCGGGTGATGAGCTACCTGAACGCCTCGTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 119 hIL12AB_019TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAGCAGCTTGT 3′UTR)CATCTCCTGGTTTTCTCTTGTCTTCCTGGCCTCGCCGCTGGTGGCCATCTGGGAGCTGAAGAAAGACGTTTACGTAGTAGAGTTGGATTGGTACCCAGACGCACCTGGAGAAATGGTGGTTCTCACCTGTGACACTCCTGAAGAAGACGGTATCACCTGGACGCTGGACCAAAGCTCAGAAGTTCTTGGCAGTGGAAAAACGCTGACCATACAAGTAAAAGAATTTGGGGATGCTGGCCAGTACACGTGCCACAAAGGAGGAGAAGTTCTCAGCCACAGTTTACTTCTTCTTCACAAGAAAGAAGATGGCATCTGGTCCACAGATATTTTAAAAGACCAGAAGGAGCCCAAGAACAAAACCTTCCTCCGCTGTGAGGCCAAGAACTACAGTGGTCGTTTCACCTGCTGGTGGCTCACCACCATCTCCACTGACCTCACCTTCTCTGTAAAAAGCAGCCGTGGTTCTTCTGACCCCCAAGGAGTCACCTGTGGGGCTGCCACGCTCTCGGCAGAAAGAGTTCGAGGTGACAACAAGGAATATGAATATTCTGTGGAATGTCAAGAAGATTCTGCCTGCCCGGCGGCAGAAGAAAGTCTTCCCATAGAAGTCATGGTGGATGCTGTTCACAAATTAAAATATGAAAACTACACCAGCAGCTTCTTCATTCGTGACATCATCAAACCAGACCCGCCCAAGAACCTTCAGTTAAAACCTTTAAAAAACAGCCGGCAGGTAGAAGTTTCCTGGGAGTACCCAGATACGTGGTCCACGCCGCACTCCTACTTCAGTTTAACCTTCTGTGTACAAGTACAAGGAAAATCAAAAAGAGAGAAGAAAGATCGTGTCTTCACTGACAAAACATCTGCCACGGTCATCTGCAGGAAGAATGCCTCCATCTCGGTTCGAGCCCAGGACCGCTACTACAGCAGCAGCTGGAGTGAGTGGGCATCTGTTCCCTGCAGTGGTGGCGGCGGCGGCGGCAGCCGCAACCTTCCTGTGGCCACGCCGGACCCTGGCATGTTCCCGTGCCTTCACCACTCCCAAAATCTTCTTCGTGCTGTTTCTAACATGCTGCAGAAGGCGCGCCAAACTTTAGAATTCTACCCGTGCACTTCTGAAGAAATAGACCATGAAGATATCACCAAAGATAAAACCAGCACGGTGGAGGCCTGCCTTCCTTTAGAGCTGACCAAGAATGAATCCTGCCTCAACAGCAGAGAGACCAGCTTCATCACCAATGGCAGCTGCCTGGCCTCGCGCAAGACCAGCTTCATGATGGCGCTGTGCCTTTCTTCCATCTATGAAGATTTAAAGATGTACCAAGTAGAATTTAAAACCATGAATGCCAAATTATTAATGGACCCCAAACGGCAGATATTTTTGGATCAAAACATGCTGGCTGTCATTGATGAGCTCATGCAAGCATTAAACTTCAACTCAGAAACTGTTCCCCAGAAGTCATCTTTAGAAGAGCCAGATTTCTACAAAACAAAAATAAAACTCTGCATTCTTCTTCATGCCTTCCGCATCCGTGCTGTCACCATTGACCGTGTCATGTCCTACTTAAATGCTTCTTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 120 hIL12AB_020TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAGCAGCTGGT 3′UTR)GATCAGCTGGTTCAGCCTGGTGTTCCTGGCTAGCCCTCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGTTGGATTGGTACCCCGACGCTCCCGGCGAGATGGTGGTGCTGACCTGCGACACCCCCGAGGAGGACGGGATCACCTGGACCCTGGATCAGTCAAGCGAGGTGCTGGGAAGCGGCAAGACCCTGACCATCCAGGTGAAGGAGTTCGGCGACGCCGGCCAATACACTTGCCACAAGGGAGGCGAGGTGCTGTCCCACTCCCTCCTGCTGCTGCACAAAAAGGAAGACGGCATCTGGAGCACCGACATCCTGAAAGACCAGAAGGAGCCTAAGAACAAAACATTCCTCAGATGCGAGGCCAAGAATTACTCCGGGAGATTCACCTGTTGGTGGCTGACCACCATCAGCACAGACCTGACCTTCAGCGTGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGTGACCTGTGGCGCCGCCACCCTGAGCGCCGAAAGAGTGCGCGGCGACAACAAGGAGTACGAGTACTCCGTGGAATGCCAGGAAGATAGCGCCTGCCCCGCCGCCGAGGAGAGCCTGCCCATCGAGGTGATGGTGGACGCCGTCCACAAGCTGAAGTACGAGAACTACACCTCTAGCTTCTTCATCAGAGATATCATCAAGCCCGATCCCCCCAAGAACCTGCAGCTGAAACCCCTGAAGAACAGCCGGCAGGTGGAGGTGAGCTGGGAGTATCCCGACACCTGGTCCACCCCCCACAGCTATTTTAGCCTGACCTTCTGCGTGCAAGTGCAGGGCAAGAGCAAGAGAGAGAAGAAGGACCGCGTGTTCACCGACAAAACCAGCGCCACCGTGATCTGCAGAAAGAACGCCAGCATCAGCGTGAGGGCCCAGGATAGATACTACAGTTCCAGCTGGAGCGAGTGGGCCAGCGTGCCCTGCAGCGGCGGCGGCGGGGGAGGCTCGAGAAACCTGCCCGTGGCTACCCCCGATCCCGGAATGTTCCCCTGCCTGCACCACAGCCAGAACCTGCTGAGGGCGGTGTCCAACATGCTTCAGAAGGCCCGGCAGACCCTGGAGTTCTACCCCTGTACCTCTGAGGAGATCGATCATGAAGATATCACAAAAGATAAAACCAGCACCGTGGAGGCCTGCCTGCCCCTGGAGCTGACCAAGAACGAGAGCTGCCTGAACTCCCGCGAGACCAGCTTCATCACGAACGGCAGCTGCCTGGCCAGCAGGAAGACCTCCTTCATGATGGCCCTGTGCCTGAGCAGCATCTACGAGGACCTGAAAATGTACCAGGTGGAGTTTAAGACCATGAACGCCAAGCTGCTGATGGACCCCAAGCGGCAAATCTTCCTGGACCAGAACATGCTGGCAGTGATCGACGAGCTCATGCAGGCCCTGAACTTCAATAGCGAGACGGTCCCCCAGAAGAGCAGCCTGGAGGAGCCCGACTTTTACAAGACCAAGATCAAGCTGTGCATCCTGCTGCACGCCTTTAGAATCCGTGCCGTGACCATTGACAGAGTGATGAGCTACCTGAATGCCAGCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 121 hIL12AB_021TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAGCAGCTGGT 3′UTR)GATCAGCTGGTTCAGCCTGGTGTTCCTGGCCAGCCCTCTGGTTGCCATCTGGGAGCTGAAGAAAGACGTGTACGTCGTGGAACTGGACTGGTATCCGGACGCCCCGGGCGAGATGGTGGTGCTGACCTGTGACACCCCCGAGGAGGACGGCATCACCTGGACGCTGGACCAATCCTCCGAGGTGCTGGGAAGCGGCAAGACCCTGACCATCCAGGTGAAGGAATTCGGGGACGCCGGGCAGTACACCTGCCACAAGGGGGGCGAAGTGCTGTCCCACTCGCTGCTGCTCCTGCATAAGAAGGAGGATGGAATCTGGTCCACCGACATCCTCAAAGATCAGAAGGAGCCCAAGAACAAGACGTTCCTGCGCTGTGAAGCCAAGAATTATTCGGGGCGATTCACGTGCTGGTGGCTGACAACCATCAGCACCGACCTGACGTTTAGCGTGAAGAGCAGCAGGGGGTCCAGCGACCCCCAGGGCGTGACGTGCGGCGCCGCCACCCTCTCCGCCGAGAGGGTGCGGGGGGACAATAAGGAGTACGAGTACAGCGTGGAATGCCAGGAGGACAGCGCCTGCCCCGCCGCGGAGGAAAGCCTCCCGATAGAGGTGATGGTGGACGCCGTGCACAAGCTCAAGTATGAGAATTACACCAGCAGCTTTTTCATCCGGGACATTATCAAGCCCGACCCCCCGAAGAACCTCCAGCTGAAGCCCCTGAAGAACAGCCGGCAGGTGGAAGTCTCCTGGGAGTATCCCGACACCTGGAGCACCCCGCACAGCTACTTCTCCCTGACCTTCTGTGTGCAGGTGCAGGGCAAGTCCAAGAGGGAAAAGAAGGACAGGGTTTTCACCGACAAGACCAGCGCGACCGTGATCTGCCGGAAGAACGCCAGCATAAGCGTCCGCGCCCAAGATAGGTACTACAGCAGCTCCTGGAGCGAGTGGGCTAGCGTGCCCTGCAGCGGGGGCGGGGGTGGGGGCTCCAGGAACCTGCCAGTGGCGACCCCCGACCCCGGCATGTTCCCCTGCCTCCATCACAGCCAGAACCTGCTGAGGGCCGTCAGCAATATGCTGCAGAAGGCCAGGCAGACCCTGGAATTCTACCCCTGCACGTCGGAGGAGATCGATCACGAGGATATCACAAAAGACAAGACTTCCACCGTGGAGGCCTGCCTGCCCCTGGAGCTCACCAAGAATGAGTCCTGTCTGAACTCCCGGGAAACCAGCTTCATCACCAACGGGTCCTGCCTGGCCAGCAGGAAGACCAGCTTTATGATGGCCCTGTGCCTGTCGAGCATCTACGAGGACCTGAAGATGTACCAGGTCGAGTTCAAGACAATGAACGCCAAGCTGCTGATGGACCCCAAGAGGCAAATCTTCCTGGACCAGAATATGCTTGCCGTCATCGACGAGCTCATGCAGGCCCTGAACTTCAACTCCGAGACCGTGCCCCAGAAGAGCAGCCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATCCTGCTGCACGCGTTCAGGATCCGGGCAGTCACCATCGACCGTGTGATGTCCTACCTGAACGCCAGCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 122 hIL12AB_022TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCATCAGCAGCTGGT 3′UTR)GATCAGCTGGTTCAGCCTGGTGTTCCTCGCCTCTCCCCTGGTGGCCATCTGGGAGCTCAAAAAGGACGTGTACGTGGTGGAGCTCGACTGGTACCCAGACGCCCCCGGGGAGATGGTGGTGCTGACCTGCGACACCCCCGAAGAAGACGGCATCACGTGGACCCTCGACCAGTCCAGCGAGGTGCTGGGGAGCGGGAAGACTCTGACCATCCAGGTCAAGGAGTTCGGGGACGCCGGGCAGTACACGTGCCACAAGGGCGGCGAAGTCTTAAGCCACAGCCTGCTCCTGCTGCACAAGAAGGAGGACGGGATCTGGTCCACAGACATACTGAAGGACCAGAAGGAGCCGAAGAATAAAACCTTTCTGAGGTGCGAGGCCAAGAACTATTCCGGCAGGTTCACGTGCTGGTGGCTTACAACAATCAGCACAGACCTGACGTTCAGCGTGAAGTCCAGCCGCGGCAGCAGCGACCCCCAGGGGGTGACCTGCGGCGCCGCCACCCTGAGCGCCGAGCGGGTGCGCGGGGACAACAAGGAGTACGAGTACTCCGTGGAGTGCCAGGAAGACAGCGCCTGTCCCGCCGCCGAAGAGAGCCTGCCTATCGAGGTCATGGTAGATGCAGTGCATAAGCTGAAGTACGAGAACTATACGAGCAGCTTTTTCATACGCGACATCATCAAGCCCGACCCCCCCAAGAACCTGCAGCTTAAGCCCCTGAAGAATAGCCGGCAGGTGGAGGTCTCCTGGGAGTACCCCGACACCTGGTCAACGCCCCACAGCTACTTCTCCCTGACCTTTTGTGTCCAAGTCCAGGGAAAGAGCAAGAGGGAGAAGAAAGATCGGGTGTTCACCGACAAGACCTCCGCCACGGTGATCTGCAGGAAGAACGCCAGCATCTCCGTGAGGGCGCAAGACAGGTACTACTCCAGCAGCTGGTCCGAATGGGCCAGCGTGCCCTGCTCCGGCGGCGGGGGCGGCGGCAGCCGAAACCTACCCGTGGCCACGCCGGATCCCGGCATGTTTCCCTGCCTGCACCACAGCCAGAACCTCCTGAGGGCCGTGTCCAACATGCTGCAGAAGGCCAGGCAGACTCTGGAGTTCTACCCCTGCACGAGCGAGGAGATCGATCACGAGGACATCACCAAGGATAAGACCAGCACTGTGGAGGCCTGCCTTCCCCTGGAGCTGACCAAGAACGAGAGCTGTCTGAACTCCAGGGAGACCTCATTCATCACCAACGGCTCCTGCCTGGCCAGCAGGAAAACCAGCTTCATGATGGCCTTGTGTCTCAGCTCCATCTACGAGGACCTGAAGATGTATCAGGTCGAGTTCAAGACAATGAACGCCAAGCTGCTGATGGACCCCAAAAGGCAGATCTTCCTGGACCAGAACATGCTGGCCGTCATCGACGAGCTGATGCAGGCCCTGAACTTCAACAGCGAGACGGTGCCCCAGAAAAGCTCCCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATCCTGCTGCACGCCTTCAGGATCAGGGCAGTGACCATCGACCGGGTGATGTCATACCTTAACGCCAGCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 123 hIL12AB_023TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCATCAGCAGCTGGT 3′UTR)GATCTCCTGGTTCAGCCTGGTGTTTCTGGCCTCGCCCCTGGTCGCCATCTGGGAGCTGAAGAAAGACGTGTACGTCGTCGAACTGGACTGGTACCCCGACGCCCCCGGGGAGATGGTGGTGCTGACCTGCGACACGCCGGAGGAGGACGGCATCACCTGGACCCTGGATCAAAGCAGCGAGGTGCTGGGCAGCGGCAAGACCCTGACCATCCAAGTGAAGGAATTCGGCGATGCCGGCCAGTACACCTGTCACAAAGGGGGCGAGGTGCTCAGCCACAGCCTGCTGCTGCTGCACAAGAAGGAGGATGGCATCTGGAGCACCGATATCCTGAAGGACCAGAAAGAGCCCAAGAACAAGACGTTCCTGAGGTGCGAGGCCAAGAACTACAGCGGTAGGTTCACGTGTTGGTGGCTGACCACCATCAGCACCGACCTGACGTTCAGCGTGAAGAGCTCCAGGGGCAGCTCCGACCCACAGGGGGTGACGTGCGGGGCCGCAACCCTCAGCGCCGAAAGGGTGCGGGGGGACAACAAGGAGTACGAATACTCCGTGGAGTGCCAGGAAGATTCGGCCTGCCCCGCCGCGGAGGAGAGCCTCCCCATCGAGGTAATGGTGGACGCCGTGCATAAGCTGAAGTACGAGAACTACACCAGCTCGTTCTTCATCCGAGACATCATCAAACCCGACCCGCCCAAAAATCTGCAGCTCAAGCCCCTGAAGAACTCCAGGCAGGTGGAGGTGAGCTGGGAGTACCCCGACACCTGGTCCACCCCGCACAGCTACTTCTCCCTGACATTCTGCGTGCAGGTGCAGGGCAAGAGCAAGCGGGAGAAGAAGGACAGGGTGTTCACCGACAAGACGAGCGCCACCGTGATCTGCCGAAAGAACGCCAGCATCTCGGTGCGCGCCCAGGATAGGTACTATTCCAGCTCCTGGAGCGAGTGGGCCTCGGTACCCTGCAGCGGCGGCGGGGGCGGCGGCAGTAGGAATCTGCCCGTGGCTACCCCGGACCCGGGCATGTTCCCCTGCCTCCACCACAGCCAGAACCTGCTGAGGGCCGTGAGCAACATGCTGCAGAAGGCCAGACAGACGCTGGAGTTCTACCCCTGCACGAGCGAGGAGATCGACCACGAGGACATCACCAAGGATAAAACTTCCACCGTCGAGGCCTGCCTGCCCTTGGAGCTGACCAAGAATGAATCCTGTCTGAACAGCAGGGAGACCTCGTTTATCACCAATGGCAGCTGCCTCGCCTCCAGGAAGACCAGCTTCATGATGGCCCTCTGTCTGAGCTCCATCTATGAGGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAACGCGAAGCTGCTGATGGACCCCAAGAGGCAGATCTTCCTGGATCAGAATATGCTGGCGGTGATCGACGAGCTCATGCAGGCCCTCAATTTCAATAGCGAGACAGTGCCCCAGAAGTCCTCCCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGTATCCTGCTGCACGCCTTCCGGATCCGGGCCGTCACCATCGACCGGGTCATGAGCTACCTCAATGCCAGCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 124 hIL12AB_024TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAGCAGCTGGT 3′UTR)GATCTCCTGGTTCTCCCTGGTGTTCCTGGCCTCGCCCCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTCGTGGAGCTCGACTGGTACCCCGACGCCCCTGGCGAGATGGTGGTGCTGACCTGCGACACCCCAGAGGAGGATGGCATCACCTGGACCCTGGATCAGTCCTCCGAGGTGCTGGGCTCCGGCAAGACGCTGACCATCCAAGTGAAGGAGTTCGGTGACGCCGGACAGTATACCTGCCATAAGGGCGGCGAGGTCCTGTCCCACAGCCTCCTCCTCCTGCATAAGAAGGAGGACGGCATCTGGAGCACCGACATCCTGAAGGACCAGAAGGAGCCCAAGAACAAGACCTTTCTGAGGTGCGAGGCCAAGAACTACAGCGGCCGATTCACCTGCTGGTGGCTCACCACCATATCCACCGACCTGACTTTCTCCGTCAAGTCCTCCCGGGGGTCCAGCGACCCCCAGGGAGTGACCTGCGGCGCCGCCACCCTCAGCGCCGAGCGGGTGCGGGGGGACAACAAGGAGTACGAATACTCCGTCGAGTGCCAGGAGGACTCCGCCTGCCCGGCCGCCGAGGAGAGCCTGCCCATCGAGGTGATGGTCGACGCGGTGCACAAGCTGAAGTACGAGAACTACACCAGCAGTTTCTTCATCAGGGATATCATCAAGCCAGATCCCCCGAAGAATCTGCAACTGAAGCCGCTGAAAAACTCACGACAGGTGGAGGTGAGCTGGGAGTACCCCGACACGTGGAGCACCCCACATTCCTACTTCAGCCTGACCTTCTGCGTGCAGGTCCAGGGCAAGAGCAAGCGGGAGAAGAAGGACAGGGTGTTCACGGATAAGACCAGTGCCACCGTGATCTGCAGGAAGAACGCCTCTATTAGCGTGAGGGCCCAGGATCGGTATTACTCCTCGAGCTGGAGCGAATGGGCCTCCGTGCCCTGCAGTGGGGGGGGTGGAGGCGGGAGCAGGAACCTGCCCGTAGCAACCCCCGACCCCGGGATGTTCCCCTGTCTGCACCACTCGCAGAACCTGCTGCGCGCGGTGAGCAACATGCTCCAAAAAGCCCGTCAGACCTTAGAGTTCTACCCCTGCACCAGCGAAGAAATCGACCACGAAGACATCACCAAGGACAAAACCAGCACCGTGGAGGCGTGCCTGCCGCTGGAGCTGACCAAGAACGAGAGCTGCCTCAACTCCAGGGAGACCAGCTTTATCACCAACGGCTCGTGCCTAGCCAGCCGGAAAACCAGCTTCATGATGGCCCTGTGCCTGAGCTCCATTTACGAGGACCTGAAGATGTATCAGGTGGAGTTCAAGACCATGAATGCCAAACTCCTGATGGACCCCAAGAGGCAGATCTTCCTGGACCAGAACATGCTCGCGGTGATCGATGAGCTGATGCAGGCCCTGAACTTTAATAGCGAGACCGTGCCCCAGAAAAGCAGCCTGGAGGAGCCGGACTTCTACAAGACCAAAATCAAGCTGTGCATCCTGCTCCACGCCTTCCGCATCCGGGCCGTGACCATCGACAGGGTGATGAGCTACCTGAACGCCAGCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 125 hIL12AB_025TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCATCAGCAGCTGGT 3′UTR)GATTTCCTGGTTCTCCCTGGTGTTCCTGGCCAGCCCCCTCGTGGCGATCTGGGAGCTAAAGAAGGACGTGTACGTGGTGGAGCTGGACTGGTACCCGGACGCACCCGGCGAGATGGTCGTTCTGACCTGCGATACGCCAGAGGAGGACGGCATCACCTGGACCCTCGATCAGAGCAGCGAGGTCCTGGGGAGCGGAAAGACCCTGACCATCCAGGTCAAGGAGTTCGGCGACGCCGGCCAGTACACCTGCCACAAAGGTGGCGAGGTCCTGAGCCACTCGCTGCTGCTCCTGCATAAGAAGGAGGACGGAATCTGGAGCACAGACATCCTGAAAGACCAGAAGGAGCCCAAGAACAAGACCTTCCTGAGGTGCGAGGCCAAGAACTACAGCGGGCGCTTCACGTGCTGGTGGCTGACCACCATCAGCACGGACCTCACCTTCTCCGTGAAGAGCAGCCGGGGATCCAGCGATCCCCAAGGCGTCACCTGCGGCGCGGCCACCCTGAGCGCGGAGAGGGTCAGGGGCGATAATAAGGAGTATGAGTACAGCGTGGAGTGCCAGGAGGACAGCGCCTGCCCGGCCGCCGAGGAGTCCCTGCCAATCGAAGTGATGGTCGACGCCGTGCACAAGCTGAAGTACGAGAACTACACCAGCAGCTTCTTCATCCGGGATATCATCAAGCCCGATCCCCCGAAGAACCTGCAGCTGAAGCCCCTCAAGAACAGCCGGCAGGTGGAGGTGAGTTGGGAGTACCCCGACACCTGGTCAACGCCCCACAGCTACTTCTCCCTGACCTTCTGTGTGCAGGTGCAGGGAAAGAGCAAGAGGGAGAAGAAAGACCGGGTCTTCACCGACAAGACCAGCGCCACGGTGATCTGCAGGAAGAACGCAAGCATCTCCGTGAGGGCCCAGGACAGGTACTACAGCTCCAGCTGGTCCGAATGGGCCAGCGTGCCCTGTAGCGGCGGCGGGGGCGGTGGCAGCCGCAACCTCCCAGTGGCCACCCCCGACCCCGGCATGTTCCCCTGCCTGCACCACAGCCAGAATCTGCTGAGGGCCGTGAGTAACATGCTGCAGAAGGCAAGGCAAACCCTCGAATTCTATCCCTGCACCTCCGAGGAGATCGACCACGAGGATATCACCAAGGACAAGACCAGCACCGTCGAGGCCTGTCTCCCCCTGGAGCTGACCAAGAATGAGAGCTGCCTGAACAGCCGGGAGACCAGCTTCATCACCAACGGGAGCTGCCTGGCCTCCAGGAAGACCTCGTTCATGATGGCGCTGTGCCTCTCAAGCATATACGAGGATCTGAAGATGTACCAGGTGGAGTTTAAGACGATGAACGCCAAGCTGCTGATGGACCCGAAGAGGCAGATCTTCCTGGACCAGAACATGCTGGCCGTGATAGACGAGCTCATGCAGGCCCTGAACTTCAACTCCGAGACCGTGCCGCAGAAGTCATCCCTCGAGGAGCCCGACTTCTATAAGACCAAGATCAAGCTGTGCATCCTGCTCCACGCCTTCCGGATAAGGGCCGTGACGATCGACAGGGTGATGAGCTACCTTAACGCCAGCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 126 hIL12AB_026TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAGCAGCTCGT 3′UTR)GATCAGCTGGTTCTCCCTGGTGTTTCTCGCCAGCCCCCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGCTGGACTGGTACCCTGACGCCCCGGGGGAGATGGTCGTGCTGACCTGCGACACCCCCGAAGAGGACGGTATCACCTGGACCCTGGACCAGTCCAGCGAGGTGCTGGGCAGCGGCAAGACCCTGACTATTCAAGTCAAGGAGTTCGGAGACGCCGGCCAGTACACCTGCCACAAGGGTGGAGAGGTGTTATCACACAGCCTGCTGCTGCTGCACAAGAAGGAAGACGGGATCTGGAGCACCGACATCCTGAAGGACCAGAAGGAGCCCAAAAACAAGACCTTCCTGCGGTGCGAGGCCAAGAACTATTCGGGCCGCTTTACGTGCTGGTGGCTGACCACCATCAGCACTGATCTCACCTTCAGCGTGAAGTCCTCCCGGGGGTCGTCCGACCCCCAGGGGGTGACCTGCGGGGCCGCCACCCTGTCCGCCGAGAGAGTGAGGGGCGATAATAAGGAGTACGAGTACAGCGTTGAGTGCCAGGAAGATAGCGCCTGTCCCGCCGCCGAGGAGAGCCTGCCCATCGAGGTGATGGTGGACGCCGTCCACAAGCTGAAGTATGAGAACTACACCTCAAGCTTCTTCATCAGGGACATCATCAAACCCGATCCGCCCAAGAATCTGCAGCTGAAGCCCCTGAAAAATAGCAGGCAGGTGGAGGTGAGCTGGGAGTACCCCGACACCTGGTCCACCCCCCATAGCTATTTCTCCCTGACGTTCTGCGTGCAGGTGCAAGGGAAGAGCAAGCGGGAGAAGAAGGACCGGGTGTTCACCGACAAGACCTCCGCCACCGTGATCTGTAGGAAGAACGCGTCGATCTCGGTCAGGGCCCAGGACAGGTATTACAGCAGCAGCTGGAGCGAGTGGGCGAGCGTGCCCTGCTCGGGCGGCGGCGGCGGCGGGAGCAGAAATCTGCCCGTGGCCACCCCAGACCCCGGAATGTTCCCCTGCCTGCACCATTCGCAGAACCTCCTGAGGGCCGTGAGCAACATGCTGCAGAAGGCCCGCCAGACGCTGGAGTTCTACCCCTGCACGAGCGAGGAGATCGACCACGAAGACATCACCAAGGACAAAACCAGCACCGTGGAGGCCTGCCTGCCCCTGGAGCTGACCAAAAACGAATCCTGCCTCAACAGCCGGGAGACCAGCTTCATCACCAACGGCAGCTGCCTGGCCAGCCGAAAGACCTCCTTCATGATGGCCCTCTGCCTGAGCAGCATCTATGAGGATCTGAAGATGTATCAGGTGGAGTTCAAGACCATGAATGCCAAGCTGCTGATGGACCCCAAGAGGCAGATATTCCTGGACCAGAATATGCTGGCCGTGATCGACGAGCTGATGCAGGCCCTGAACTTCAACAGCGAGACCGTCCCCCAGAAGTCCAGCCTGGAGGAGCCGGACTTTTACAAAACGAAGATCAAGCTGTGCATACTGCTGCACGCCTTCAGGATCCGGGCCGTGACAATCGACAGGGTGATGTCCTACCTGAACGCCAGCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 127 hIL12AB_027TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGTCACCAGCAGCTGGT 3′UTR)GATCAGCTGGTTCTCCCTGGTGTTCCTGGCCAGCCCCCTGGTGGCCATCTGGGAGCTCAAGAAGGACGTCTACGTCGTGGAGCTGGATTGGTACCCCGACGCTCCCGGGGAGATGGTGGTGCTGACCTGCGACACCCCCGAGGAGGACGGCATCACCTGGACGCTGGACCAGAGCTCAGAGGTGCTGGGAAGCGGAAAGACACTGACCATCCAGGTGAAGGAGTTCGGGGATGCCGGGCAGTATACCTGCCACAAGGGCGGCGAAGTGCTGAGCCATTCCCTGCTGCTGCTGCACAAGAAGGAGGACGGCATATGGTCCACCGACATCCTGAAGGATCAGAAGGAGCCGAAGAATAAAACCTTCCTGAGGTGCGAGGCCAAGAATTACAGCGGCCGATTCACCTGCTGGTGGCTGACCACCATCAGCACCGACCTGACCTTCAGTGTGAAGTCCTCACGGGGCAGCTCAGATCCCCAGGGCGTGACCTGCGGGGCCGCGACACTCAGCGCCGAGCGGGTGAGGGGTGATAACAAGGAGTACGAGTATTCTGTGGAGTGCCAGGAAGACTCCGCCTGTCCCGCCGCCGAGGAGTCCCTGCCCATCGAGGTGATGGTGGACGCCGTGCATAAACTGAAGTACGAGAACTACACCTCCAGCTTCTTCATCCGGGATATAATCAAGCCCGACCCTCCGAAAAACCTGCAGCTGAAGCCCCTTAAAAACAGCCGGCAGGTGGAGGTGAGCTGGGAGTACCCCGACACCTGGAGCACCCCCCATAGCTATTTCAGCCTGACCTTCTGCGTGCAGGTGCAGGGGAAGTCCAAGCGCGAGAAAAAGGACCGGGTGTTCACCGACAAGACGAGCGCCACCGTGATCTGCCGGAAGAACGCCAGTATAAGCGTAAGGGCCCAGGATAGGTACTACAGCTCCAGCTGGTCGGAGTGGGCCTCCGTGCCCTGTTCCGGCGGCGGGGGGGGTGGCAGCAGGAACCTCCCCGTGGCCACGCCGGACCCCGGCATGTTCCCGTGCCTGCACCACTCCCAAAACCTCCTGCGGGCCGTCAGCAACATGCTGCAAAAGGCGCGGCAGACCCTGGAGTTTTACCCCTGTACCTCCGAAGAGATCGACCACGAGGATATCACCAAGGATAAGACCTCCACCGTGGAGGCCTGTCTCCCCCTGGAGCTGACCAAGAACGAGAGCTGTCTTAACAGCAGAGAGACCTCGTTCATAACGAACGGCTCCTGCCTCGCTTCCAGGAAGACGTCGTTCATGATGGCGCTGTGCCTGTCCAGCATCTACGAGGACCTGAAGATGTATCAGGTCGAGTTCAAAACCATGAACGCCAAGCTGCTGATGGACCCCAAGAGGCAGATCTTCCTGGACCAGAACATGCTCGCCGTGATCGACGAGCTGATGCAGGCCCTGAACTTCAACAGCGAAACCGTGCCCCAGAAGTCAAGCCTGGAGGAGCCGGACTTCTATAAGACCAAGATCAAGCTGTGTATCCTGCTACACGCTTTTCGTATCCGGGCCGTGACCATCGACAGGGTTATGTCGTACTTGAACGCCAGCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 128 hIL12AB_028TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAACAGCTCGT 3′UTR)GATCAGCTGGTTCAGCCTGGTGTTCCTGGCCAGCCCGCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGCTGGACTGGTACCCCGACGCCCCCGGCGAGATGGTGGTCCTGACCTGCGACACGCCGGAAGAGGACGGCATCACCTGGACCCTGGATCAGTCCAGCGAGGTGCTGGGCTCCGGCAAGACCCTGACCATTCAGGTGAAGGAGTTCGGCGACGCCGGTCAGTACACCTGCCACAAGGGCGGCGAGGTGCTGAGCCACAGCCTACTGCTCCTGCACAAAAAGGAGGATGGAATCTGGTCCACCGACATCCTCAAGGACCAGAAGGAGCCGAAGAACAAGACGTTCCTCCGGTGCGAGGCCAAGAACTACAGCGGCAGGTTTACCTGCTGGTGGCTGACCACCATCAGCACCGACCTGACATTTTCCGTGAAGAGCAGCCGCGGCAGCAGCGATCCCCAGGGCGTGACCTGCGGGGCGGCCACCCTGTCCGCCGAGCGTGTGAGGGGCGACAACAAGGAGTACGAGTACAGCGTGGAATGCCAGGAGGACAGCGCCTGTCCCGCCGCCGAGGAGAGCCTGCCAATCGAGGTCATGGTGGACGCCGTGCACAAGCTGAAGTACGAGAACTACACGAGCAGCTTCTTCATCAGGGACATCATCAAACCGGACCCGCCCAAGAACCTGCAGCTGAAACCCTTGAAAAACAGCAGGCAGGTGGAAGTGTCTTGGGAGTACCCCGACACCTGGTCCACCCCCCACAGCTACTTTAGCCTGACCTTCTGTGTGCAGGTCCAGGGCAAGTCCAAGAGGGAGAAGAAGGACAGGGTGTTCACCGACAAAACCAGCGCCACCGTGATCTGCAGGAAGAACGCCTCCATCAGCGTGCGGGCCCAGGACAGGTATTACAGCTCGTCGTGGAGCGAGTGGGCCAGCGTGCCCTGCTCCGGGGGAGGCGGCGGCGGAAGCCGGAATCTGCCCGTGGCCACCCCCGATCCCGGCATGTTCCCGTGTCTGCACCACAGCCAGAACCTGCTGCGGGCCGTGAGCAACATGCTGCAGAAGGCCCGCCAAACCCTGGAGTTCTACCCCTGTACAAGCGAGGAGATCGACCATGAGGACATTACCAAGGACAAGACCAGCACCGTGGAGGCCTGCCTGCCCCTCGAGCTCACAAAGAACGAATCCTGCCTGAATAGCCGCGAGACCAGCTTTATCACGAACGGGTCCTGCCTCGCCAGCCGGAAGACAAGCTTCATGATGGCCCTGTGCCTGAGCAGCATCTACGAGGACCTGAAAATGTACCAAGTGGAGTTCAAAACGATGAACGCCAAGCTGCTGATGGACCCCAAGCGCCAGATCTTCCTGGACCAGAACATGCTGGCCGTCATCGACGAGCTCATGCAGGCCCTGAACTTCAACAGCGAGACCGTGCCCCAGAAGAGCAGCCTGGAGGAGCCCGACTTCTACAAGACGAAGATCAAGCTCTGCATCCTGCTGCACGCTTTCCGCATCCGCGCGGTGACCATCGACCGGGTGATGAGCTACCTCAACGCCAGTTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 129 hIL12AB_029TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAACAGCTGGT 3′UTR)GATCAGCTGGTTCAGCCTGGTGTTTCTGGCCTCCCCTCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGCTGGACTGGTACCCTGACGCCCCCGGCGAAATGGTGGTGCTGACGTGCGACACCCCCGAGGAGGATGGCATCACCTGGACCCTGGACCAAAGCAGCGAGGTCCTCGGAAGCGGCAAGACCCTCACTATCCAAGTGAAGGAGTTCGGGGATGCGGGCCAGTACACCTGCCACAAGGGCGGCGAGGTGCTGTCTCATAGCCTGCTGCTCCTGCATAAGAAGGAAGACGGCATCTGGAGCACCGACATACTGAAGGATCAGAAGGAGCCCAAGAACAAGACCTTCCTGAGGTGCGAGGCCAAGAACTACTCCGGGCGCTTCACCTGTTGGTGGCTGACCACCATCTCCACCGACCTGACCTTCAGCGTGAAGAGCAGCAGGGGGAGCAGCGACCCCCAGGGGGTGACCTGCGGAGCCGCGACCTTGTCGGCCGAGCGGGTGAGGGGCGACAATAAGGAGTACGAGTACTCGGTCGAATGCCAGGAGGACTCCGCCTGCCCCGCCGCCGAGGAGTCCCTCCCCATCGAAGTGATGGTGGACGCCGTCCACAAGCTGAAGTACGAGAACTACACCAGCAGCTTCTTCATACGGGATATCATCAAGCCCGACCCCCCGAAGAACCTGCAGCTGAAACCCTTGAAGAACTCCAGGCAGGTGGAGGTGAGCTGGGAGTACCCCGACACCTGGTCCACCCCGCACTCATACTTCAGCCTGACCTTCTGTGTACAGGTCCAGGGCAAGAGCAAGAGGGAAAAGAAGGATAGGGTGTTCACCGACAAGACCTCCGCCACGGTGATCTGTCGGAAAAACGCCAGCATCTCCGTGCGGGCCCAGGACAGGTACTATTCCAGCAGCTGGAGCGAGTGGGCCTCCGTCCCCTGCTCCGGCGGCGGTGGCGGGGGCAGCAGGAACCTCCCCGTGGCCACCCCCGATCCCGGGATGTTCCCATGCCTGCACCACAGCCAAAACCTGCTGAGGGCCGTCTCCAATATGCTGCAGAAGGCGAGGCAGACCCTGGAGTTCTACCCCTGTACCTCCGAGGAGATCGACCACGAGGATATCACCAAGGACAAGACCTCCACGGTCGAGGCGTGCCTGCCCCTGGAGCTCACGAAGAACGAGAGCTGCCTTAACTCCAGGGAAACCTCGTTTATCACGAACGGCAGCTGCCTGGCGTCACGGAAGACCTCCTTTATGATGGCCCTATGTCTGTCCTCGATCTACGAGGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAACGCCAAGCTGCTGATGGATCCCAAGAGGCAGATTTTCCTGGACCAGAACATGCTGGCCGTGATTGACGAGCTGATGCAGGCGCTGAACTTCAACAGCGAGACAGTGCCGCAGAAGAGCTCCCTGGAGGAGCCGGACTTTTACAAGACCAAGATAAAGCTGTGCATCCTGCTCCACGCCTTCAGAATACGGGCCGTCACCATCGATAGGGTGATGTCTTACCTGAACGCCTCCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 130 hIL12AB_030TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAGCAGCTGGT 3′UTR)GATTAGCTGGTTTAGCCTGGTGTTCCTGGCAAGCCCCCTGGTGGCCATCTGGGAACTGAAAAAGGACGTGTACGTGGTCGAGCTGGATTGGTACCCCGACGCCCCCGGCGAAATGGTGGTGCTGACGTGTGATACCCCCGAGGAGGACGGGATCACCTGGACCCTGGATCAGAGCAGCGAGGTGCTGGGGAGCGGGAAGACCCTGACGATCCAGGTCAAGGAGTTCGGCGACGCTGGGCAGTACACCTGTCACAAGGGCGGGGAGGTGCTGTCCCACTCCCTGCTGCTCCTGCATAAGAAAGAGGACGGCATCTGGTCCACCGACATCCTCAAGGACCAGAAGGAGCCCAAGAACAAGACCTTCCTGCGGTGTGAGGCGAAGAACTACAGCGGCCGTTTCACCTGCTGGTGGCTGACGACAATCAGCACCGACTTGACGTTCTCCGTGAAGTCCTCCAGAGGCAGCTCCGACCCCCAAGGGGTGACGTGCGGCGCGGCCACCCTGAGCGCCGAGCGGGTGCGGGGGGACAACAAGGAGTACGAGTACTCCGTGGAGTGCCAGGAGGACAGCGCCTGTCCCGCAGCCGAGGAGTCCCTGCCCATCGAAGTCATGGTGGACGCCGTCCACAAGCTGAAGTACGAGAACTACACCAGCAGCTTCTTCATCCGCGATATCATCAAGCCCGATCCCCCCAAAAACCTGCAACTGAAGCCGCTGAAGAATAGCAGGCAGGTGGAGGTGTCCTGGGAGTACCCGGACACCTGGAGCACGCCCCACAGCTATTTCAGCCTGACCTTTTGCGTGCAGGTCCAGGGGAAGAGCAAGCGGGAGAAGAAGGACCGCGTGTTTACGGACAAAACCAGCGCCACCGTGATCTGCAGGAAGAACGCCAGCATCAGCGTGAGGGCCCAGGACAGGTACTACAGCAGCTCCTGGAGCGAGTGGGCCTCCGTGCCCTGTTCCGGAGGCGGCGGGGGCGGTTCCCGGAACCTCCCGGTGGCCACCCCCGACCCGGGCATGTTCCCGTGCCTGCACCACTCACAGAATCTGCTGAGGGCCGTGAGCAATATGCTGCAGAAGGCAAGGCAGACCCTGGAGTTTTATCCCTGCACCAGCGAGGAGATCGACCACGAAGACATCACCAAGGACAAGACCAGCACAGTGGAGGCCTGCCTGCCCCTGGAACTGACCAAGAACGAGTCCTGTCTGAACTCCCGGGAAACCAGCTTCATAACCAACGGCTCCTGTCTCGCCAGCAGGAAGACCAGCTTCATGATGGCCCTGTGCCTCAGCTCCATCTACGAGGACCTCAAGATGTACCAGGTTGAGTTCAAGACCATGAACGCCAAGCTCCTGATGGACCCCAAGAGGCAGATCTTCCTGGACCAGAATATGCTGGCCGTGATCGATGAGTTAATGCAGGCGCTGAACTTCAACAGCGAGACGGTGCCCCAAAAGTCCTCGCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATCCTCCTGCACGCCTTCCGAATCCGGGCCGTAACCATCGACAGGGTGATGAGCTATCTCAACGCCTCCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 131 hIL12AB_031TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAGCAGCTCGT 3′UTR)GATCAGCTGGTTCTCGCTTGTGTTCCTGGCCTCCCCCCTCGTCGCCATCTGGGAGCTGAAGAAAGACGTGTACGTGGTGGAGCTGGACTGGTATCCCGACGCCCCGGGGGAGATGGTGGTGCTGACCTGCGACACCCCGGAAGAGGACGGCATCACCTGGACGCTCGACCAGTCGTCCGAAGTGCTGGGGTCGGGCAAGACCCTCACCATCCAGGTGAAGGAGTTCGGAGACGCCGGCCAGTACACCTGTCATAAGGGGGGGGAGGTGCTGAGCCACAGCCTCCTGCTCCTGCACAAAAAGGAGGACGGCATCTGGAGCACCGATATCCTCAAGGACCAGAAGGAGCCCAAGAACAAGACGTTCCTGAGGTGTGAGGCCAAGAACTACAGCGGGCGGTTCACGTGTTGGTGGCTCACCACCATCTCCACCGACCTCACCTTCTCCGTGAAGTCAAGCAGGGGCAGCTCCGACCCCCAAGGCGTCACCTGCGGCGCCGCCACCCTGAGCGCCGAGAGGGTCAGGGGGGATAACAAGGAATACGAGTACAGTGTGGAGTGCCAAGAGGATAGCGCCTGTCCCGCCGCCGAAGAGAGCCTGCCCATCGAAGTGATGGTGGACGCCGTGCACAAGCTGAAGTACGAGAACTACACCTCCAGCTTCTTCATCAGGGATATCATCAAGCCCGATCCCCCCAAGAACCTGCAGCTGAAGCCCCTGAAGAACAGCAGGCAGGTGGAGGTGAGCTGGGAGTATCCCGACACGTGGAGCACCCCGCACAGCTACTTCTCGCTGACCTTCTGCGTGCAGGTGCAAGGGAAGTCCAAGAGGGAGAAGAAGGATAGGGTGTTCACCGACAAAACGAGCGCCACCGTGATCTGCCGGAAGAATGCCAGCATCTCTGTGAGGGCCCAGGACAGGTACTATTCCAGCTCCTGGTCGGAGTGGGCCAGCGTGCCCTGTAGCGGCGGGGGCGGGGGCGGCAGCAGGAACCTCCCGGTTGCCACCCCCGACCCCGGCATGTTTCCGTGCCTGCACCACTCGCAAAACCTGCTGCGCGCGGTCTCCAACATGCTGCAAAAAGCGCGCCAGACGCTGGAGTTCTACCCCTGCACCAGCGAGGAGATCGATCATGAAGATATCACCAAAGACAAGACCTCGACCGTGGAGGCCTGCCTGCCCCTGGAGCTCACCAAGAACGAAAGCTGCCTGAACAGCAGGGAGACAAGCTTCATCACCAACGGCAGCTGCCTGGCCTCCCGGAAGACCAGCTTCATGATGGCCCTGTGCCTGTCCAGCATCTACGAGGATCTGAAGATGTACCAAGTGGAGTTTAAGACCATGAACGCCAAGCTGTTAATGGACCCCAAAAGGCAGATCTTCCTGGATCAGAACATGCTGGCCGTCATCGACGAGCTGATGCAAGCCCTGAACTTCAACAGCGAGACGGTGCCCCAGAAGAGCAGCCTCGAGGAGCCCGACTTCTATAAGACCAAGATAAAGCTGTGCATTCTGCTGCACGCCTTCAGAATCAGGGCCGTGACCATCGATAGGGTGATGAGCTACCTGAACGCCAGCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 132 hIL12AB_032TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGTCACCAGCAGCTGGT 3′UTR)GATTTCCTGGTTCAGTCTGGTGTTTCTTGCCAGCCCCCTGGTGGCCATCTGGGAGCTGAAGAAAGACGTATACGTCGTGGAGCTGGACTGGTATCCCGACGCTCCCGGCGAGATGGTGGTCCTCACCTGCGACACCCCAGAGGAGGACGGCATCACCTGGACCCTGGACCAGAGCTCCGAGGTCCTGGGCAGCGGTAAGACCCTCACCATCCAGGTGAAGGAGTTTGGTGATGCCGGGCAGTATACCTGCCACAAGGGCGGCGAGGTGCTGTCCCACAGCCTCCTGTTACTGCATAAGAAGGAGGATGGCATCTGGAGCACCGACATCCTCAAGGACCAGAAAGAGCCCAAGAACAAGACCTTTCTGCGGTGCGAGGCGAAAAATTACTCCGGCCGGTTCACCTGCTGGTGGCTGACCACCATCAGCACGGACCTGACGTTCTCCGTGAAGTCGAGCAGGGGGAGCTCCGATCCCCAGGGCGTGACCTGCGGCGCGGCCACCCTGAGCGCCGAGCGCGTCCGCGGGGACAATAAGGAATACGAATATAGCGTGGAGTGCCAGGAGGACAGCGCCTGCCCCGCGGCCGAGGAGAGCCTCCCGATCGAGGTGATGGTGGATGCCGTCCACAAGCTCAAATACGAAAACTACACCAGCAGCTTCTTCATTAGGGACATCATCAAGCCCGACCCCCCCAAAAACCTGCAGCTGAAGCCCCTGAAGAACAGCCGCCAGGTCGAGGTGTCATGGGAGTACCCAGACACCTGGAGCACCCCCCACTCCTACTTCAGCCTGACCTTCTGCGTCCAGGTGCAGGGAAAGTCCAAACGGGAGAAGAAGGATAGGGTCTTTACCGATAAGACGTCGGCCACCGTCATCTGCAGGAAGAACGCCAGCATAAGCGTGCGGGCGCAGGATCGGTACTACAGCTCGAGCTGGTCCGAATGGGCCTCCGTGCCCTGTAGCGGAGGGGGTGGCGGGGGCAGCAGGAACCTGCCCGTGGCCACCCCGGACCCGGGCATGTTTCCCTGCCTGCATCACAGTCAGAACCTGCTGAGGGCCGTGAGCAACATGCTCCAGAAGGCCCGCCAGACCCTGGAGTTTTACCCCTGCACCAGCGAAGAGATCGATCACGAAGACATCACCAAAGACAAGACCTCCACCGTGGAGGCCTGTCTGCCCCTGGAGCTGACCAAGAACGAGAGCTGTCTGAACAGCAGGGAGACCTCCTTCATCACCAACGGCTCCTGCCTGGCATCCCGGAAGACCAGCTTCATGATGGCCCTGTGTCTGAGCTCTATCTACGAGGACCTGAAGATGTACCAGGTCGAGTTCAAGACCATGAACGCCAAGCTGCTGATGGACCCCAAGCGACAGATATTCCTGGACCAGAACATGCTCGCCGTGATCGATGAACTGATGCAAGCCCTGAACTTCAATAGCGAGACCGTGCCCCAGAAAAGCAGCCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAACTGTGCATACTGCTGCACGCGTTCAGGATCCGGGCCGTCACCATCGACCGGGTGATGTCCTATCTGAATGCCAGCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 133 hIL12AB_033TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAGCAGCTCGT 3′UTR)GATTAGCTGGTTTTCGCTGGTGTTCCTGGCCAGCCCTCTCGTGGCCATCTGGGAGCTGAAAAAAGACGTGTACGTGGTGGAGCTGGACTGGTACCCGGACGCCCCCGGCGAGATGGTGGTGCTGACGTGCGACACCCCGGAAGAGGACGGCATCACCTGGACCCTGGACCAGTCATCCGAGGTCCTGGGCAGCGGCAAGACGCTCACCATCCAGGTGAAGGAGTTCGGCGACGCCGGCCAGTACACATGCCATAAGGGCGGGGAGGTGCTGAGCCACAGCCTGCTCCTCCTGCACAAGAAGGAGGATGGCATCTGGTCTACAGACATCCTGAAGGACCAGAAAGAGCCCAAGAACAAGACCTTCCTCCGGTGCGAGGCCAAGAACTACTCCGGGCGGTTTACTTGTTGGTGGCTGACCACCATCAGCACCGACCTCACCTTCAGCGTGAAGAGCTCCCGAGGGAGCTCCGACCCCCAGGGGGTCACCTGCGGCGCCGCCACCCTGAGCGCCGAGCGGGTGAGGGGCGACAACAAGGAGTATGAATACAGCGTGGAATGCCAAGAGGACAGCGCCTGTCCCGCGGCCGAGGAAAGCCTGCCCATCGAGGTGATGGTGGACGCCGTCCACAAACTCAAGTACGAGAACTACACCAGCAGTTTCTTCATTCGCGACATCATCAAGCCGGACCCCCCCAAAAACCTGCAGCTCAAACCCCTGAAGAACAGCAGGCAGGTGGAGGTCAGCTGGGAGTACCCGGACACCTGGAGCACCCCCCATAGCTACTTCAGCCTGACCTTCTGCGTGCAGGTGCAGGGCAAGAGCAAACGCGAGAAGAAGGACCGGGTGTTTACCGACAAGACCAGCGCCACGGTGATCTGCCGAAAGAATGCAAGCATCTCCGTGAGGGCGCAGGACCGCTACTACTCTAGCAGCTGGAGCGAGTGGGCCAGCGTGCCCTGCAGCGGTGGCGGCGGAGGCGGCAGCCGTAACCTCCCCGTGGCCACCCCCGACCCCGGCATGTTCCCGTGTCTGCACCACTCCCAGAACCTGCTGAGGGCCGTCAGCAATATGCTGCAGAAGGCCCGGCAGACGCTGGAGTTCTACCCCTGCACCTCCGAGGAGATCGACCATGAGGACATTACCAAGGACAAGACGAGCACTGTGGAGGCCTGCCTGCCCCTGGAGCTCACCAAAAACGAGAGCTGCCTGAATAGCAGGGAGACGTCCTTCATCACCAACGGCAGCTGTCTGGCCAGCAGGAAGACCAGCTTCATGATGGCCCTGTGCCTCTCCTCCATATATGAGGATCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAACGCCAAGCTGCTGATGGATCCCAAGAGGCAGATCTTCCTGGACCAGAATATGCTGGCCGTGATTGACGAGCTGATGCAGGCCCTGAACTTTAATAGCGAGACCGTCCCCCAGAAGAGCAGCCTGGAGGAGCCCGACTTCTATAAGACCAAGATCAAGCTGTGCATACTGCTGCACGCGTTTAGGATAAGGGCCGTCACCATCGACAGGGTGATGAGCTACCTGAATGCCAGCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 134 hIL12AB_034TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAACAGCTGGT 3′UTR)GATCTCCTGGTTCAGCCTGGTGTTCCTCGCCAGCCCCCTGGTGGCCATCTGGGAGCTGAAGAAAGACGTGTACGTGGTGGAGCTGGACTGGTATCCCGACGCCCCCGGCGAGATGGTCGTGCTGACCTGCGACACCCCGGAGGAGGACGGCATCACCTGGACCCTGGATCAGTCCTCCGAGGTGCTGGGCAGCGGGAAGACCCTGACCATCCAGGTGAAAGAGTTCGGAGATGCCGGCCAGTATACCTGTCACAAGGGGGGTGAGGTGCTGAGCCATAGCCTCTTGCTTCTGCACAAGAAGGAGGACGGCATCTGGTCCACCGACATCCTCAAGGACCAAAAGGAGCCGAAGAATAAAACGTTCCTGAGGTGCGAAGCCAAGAACTATTCCGGACGGTTCACCTGCTGGTGGCTGACCACCATCAGCACCGACCTCACCTTCTCCGTAAAGTCAAGCAGGGGCAGCTCCGACCCCCAGGGCGTGACCTGCGGAGCCGCCACCCTGAGCGCAGAGAGGGTGAGGGGCGACAACAAGGAGTACGAATACTCCGTCGAGTGCCAGGAGGACAGCGCCTGCCCCGCCGCCGAGGAAAGTCTGCCCATCGAGGTGATGGTGGACGCCGTGCACAAGCTCAAATACGAGAACTACACCAGCAGCTTCTTCATCCGGGATATCATCAAGCCCGACCCTCCAAAGAATCTGCAGCTGAAACCCCTTAAGAACAGCAGGCAGGTGGAGGTCAGCTGGGAGTACCCCGACACCTGGAGCACGCCCCACTCCTACTTTAGCCTGACCTTTTGCGTGCAGGTGCAGGGGAAAAGCAAGCGGGAGAAGAAGGACAGGGTGTTCACCGATAAGACCTCCGCTACCGTGATCTGCAGGAAGAACGCCTCAATCAGCGTGAGGGCCCAGGATCGGTACTACTCCAGCTCCTGGAGCGAGTGGGCCAGCGTGCCCTGCTCTGGCGGTGGCGGCGGGGGCAGCCGGAACCTGCCGGTGGCCACTCCCGACCCGGGCATGTTCCCGTGCCTCCACCATTCCCAGAACCTGCTGCGGGCCGTGTCCAATATGCTCCAGAAGGCAAGGCAGACCCTGGAGTTCTACCCCTGCACCAGCGAGGAGATCGATCACGAGGACATCACCAAAGACAAAACCAGCACGGTCGAGGCCTGCCTGCCCCTGGAACTCACCAAGAACGAAAGCTGTCTCAACAGCCGCGAGACCAGCTTCATAACCAACGGTTCCTGTCTGGCCTCCCGCAAGACCAGCTTTATGATGGCCCTCTGTCTGAGCTCCATCTATGAAGACCTGAAAATGTACCAGGTGGAGTTCAAAACCATGAACGCCAAGCTTCTGATGGACCCCAAGAGGCAGATCTTCCTGGATCAGAACATGCTGGCCGTGATCGACGAGCTGATGCAGGCCCTGAACTTTAACTCCGAGACCGTGCCCCAGAAAAGCAGCCTGGAAGAGCCCGATTTCTACAAAACGAAGATCAAGCTGTGCATCCTGCTGCACGCCTTCCGGATCCGTGCGGTGACCATCGATAGGGTGATGAGCTACCTGAACGCCAGCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 135 hIL12AB_035TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAACAGCTGGT 3′UTR)AATCAGCTGGTTCAGCCTGGTTTTCCTCGCGTCGCCCCTGGTGGCCATCTGGGAGTTAAAGAAGGACGTGTACGTGGTGGAGCTGGATTGGTACCCCGACGCCCCGGGCGAGATGGTCGTGCTCACCTGCGATACCCCCGAGGAGGACGGGATCACCTGGACCCTGGACCAATCCAGCGAGGTGCTGGGCAGCGGCAAGACCCTGACCATACAGGTGAAGGAATTTGGGGACGCCGGGCAGTACACCTGCCACAAGGGCGGGGAAGTGCTGTCCCACTCCCTCCTGCTGCTGCATAAGAAGGAGGACGGCATCTGGAGCACCGACATCCTGAAGGACCAAAAGGAGCCCAAGAACAAGACCTTCCTGAGGTGCGAGGCCAAAAACTATTCCGGCCGCTTTACCTGTTGGTGGCTGACCACCATCTCCACCGATCTGACCTTCAGCGTGAAGTCGTCTAGGGGCTCCTCCGACCCCCAGGGCGTAACCTGCGGCGCCGCGACCCTGAGCGCCGAGAGGGTGCGGGGCGATAACAAAGAGTACGAGTACTCGGTGGAGTGCCAGGAGGACAGCGCCTGTCCGGCGGCCGAGGAGAGCCTGCCCATCGAGGTGATGGTGGACGCCGTCCACAAGCTGAAGTACGAGAACTACACCAGTTCGTTCTTCATCAGGGACATCATCAAGCCGGACCCCCCCAAGAACCTCCAGCTGAAGCCCCTGAAGAACAGCAGGCAGGTGGAAGTGTCCTGGGAGTATCCCGACACCTGGAGCACCCCCCACAGCTACTTCAGCCTGACCTTTTGCGTGCAGGTGCAGGGCAAAAGCAAGAGGGAAAAGAAGGACCGGGTGTTCACCGATAAGACGAGCGCCACCGTTATCTGCAGGAAGAACGCCTCCATAAGCGTGAGGGCGCAGGACCGTTACTACAGCAGCAGCTGGAGTGAGTGGGCAAGCGTGCCCTGTAGCGGCGGGGGCGGGGGCGGGTCCCGCAACCTCCCCGTCGCCACCCCCGACCCAGGCATGTTTCCGTGCCTGCACCACAGCCAGAACCTGCTGCGGGCCGTTAGCAACATGCTGCAGAAGGCCAGGCAGACCCTCGAGTTCTATCCCTGCACATCTGAGGAGATCGACCACGAAGACATCACTAAGGATAAGACCTCCACCGTGGAGGCCTGTCTGCCCCTCGAGCTGACCAAGAATGAATCCTGCCTGAACAGCCGAGAGACCAGCTTTATCACCAACGGCTCCTGCCTGGCCAGCAGGAAGACCTCCTTCATGATGGCCCTGTGCCTCTCCAGCATCTACGAGGATCTGAAGATGTACCAGGTAGAGTTCAAGACGATGAACGCCAAGCTCCTGATGGACCCCAAGAGGCAGATATTCCTGGACCAGAACATGCTGGCGGTGATCGACGAGCTGATGCAGGCCCTGAATTTCAACAGCGAGACGGTGCCACAGAAGTCCAGCCTGGAGGAGCCAGACTTCTACAAGACCAAGATCAAACTGTGCATCCTCCTGCACGCGTTCAGGATCCGCGCCGTCACCATAGACAGGGTGATGAGTTATCTGAACGCCAGCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 136 hIL12AB_036TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCATCAGCAGCTGGT 3′UTR)AATCAGCTGGTTTAGCCTGGTGTTCCTGGCCAGCCCACTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAACTGGACTGGTACCCCGACGCCCCTGGCGAGATGGTGGTACTGACCTGTGACACCCCGGAGGAAGACGGTATCACCTGGACCCTGGATCAGAGCTCCGAGGTGCTGGGCTCCGGCAAGACACTGACCATCCAAGTTAAGGAATTTGGGGACGCCGGCCAGTACACCTGCCACAAGGGGGGCGAGGTGCTGTCCCACTCCCTGCTGCTTCTGCATAAGAAGGAGGATGGCATCTGGTCCACCGACATACTGAAGGACCAGAAGGAGCCCAAGAATAAGACCTTCCTGAGATGCGAGGCCAAGAACTACTCGGGAAGGTTCACCTGCTGGTGGCTGACCACCATCAGCACCGACCTGACCTTCTCCGTGAAGAGCTCCCGGGGCAGCTCCGACCCCCAGGGCGTAACCTGTGGGGCCGCTACCCTGTCCGCCGAGAGGGTCCGGGGCGACAACAAGGAATACGAGTACAGCGTGGAGTGCCAGGAGGACTCCGCCTGCCCCGCCGCCGAGGAGTCGCTGCCCATAGAGGTGATGGTGGACGCCGTGCACAAGCTCAAGTACGAGAATTACACCAGCAGCTTCTTTATCAGGGACATAATTAAGCCGGACCCCCCAAAGAATCTGCAGCTGAAGCCCCTGAAGAATAGCCGGCAGGTGGAAGTGTCCTGGGAGTACCCCGACACCTGGAGCACCCCCCACTCCTATTTCTCACTGACATTCTGCGTGCAGGTGCAAGGGAAAAGCAAGAGGGAGAAGAAGGATAGGGTGTTCACCGACAAGACAAGCGCCACCGTGATCTGCCGAAAAAATGCCAGCATCAGCGTGAGGGCCCAGGATCGGTATTACAGCAGCTCCTGGAGCGAGTGGGCCAGCGTGCCCTGTTCCGGCGGGGGAGGGGGCGGCTCCCGGAACCTGCCGGTGGCCACCCCCGACCCTGGCATGTTCCCCTGCCTGCATCACAGCCAGAACCTGCTCCGGGCCGTGTCGAACATGCTGCAGAAGGCCCGGCAGACCCTCGAGTTTTACCCCTGCACCAGCGAAGAGATCGACCACGAAGACATAACCAAGGACAAGACCAGCACGGTGGAGGCCTGCCTGCCCCTGGAGCTTACCAAAAACGAGTCCTGCCTGAACAGCCGGGAAACCAGCTTCATAACGAACGGGAGCTGCCTGGCCTCCAGGAAGACCAGCTTCATGATGGCGCTGTGTCTGTCCAGCATATACGAGGATCTGAAGATGTATCAGGTGGAATTCAAAACTATGAATGCCAAGCTCCTGATGGACCCCAAGAGGCAGATCTTCCTGGACCAGAACATGCTAGCCGTGATCGACGAGCTGATGCAGGCCCTCAACTTCAACTCGGAGACGGTGCCCCAGAAGTCCAGCCTCGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATACTGCTGCATGCCTTCAGGATAAGGGCGGTGACTATCGACAGGGTCATGTCCTACCTGAACGCCAGCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 137 hIL12AB_037TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAACAACTGGT 3′UTR)GATCAGCTGGTTCTCCCTGGTGTTCCTGGCCAGCCCCCTGGTGGCCATCTGGGAGCTCAAAAAAGACGTGTACGTGGTGGAGCTCGATTGGTACCCAGACGCGCCGGGGGAAATGGTGGTGCTGACCTGCGACACCCCAGAGGAGGATGGCATCACGTGGACGCTGGATCAGTCCAGCGAGGTGCTGGGGAGCGGCAAGACGCTCACCATCCAGGTGAAGGAATTTGGCGACGCGGGCCAGTATACCTGTCACAAGGGCGGCGAGGTGCTGAGCCACTCCCTGCTGCTGCTGCACAAGAAGGAGGATGGGATCTGGTCAACCGATATCCTGAAAGACCAGAAGGAGCCCAAGAACAAGACCTTCCTGCGCTGCGAGGCCAAGAACTATAGCGGCAGGTTCACCTGCTGGTGGCTGACCACCATCAGCACCGACCTGACCTTCAGCGTGAAATCCTCCAGGGGCAGCAGCGACCCCCAGGGCGTGACCTGCGGTGCCGCCACGCTCTCCGCCGAGCGAGTGAGGGGTGACAACAAGGAGTACGAGTACAGCGTGGAATGTCAGGAGGACAGCGCCTGTCCCGCCGCCGAGGAGTCGCTGCCCATCGAGGTGATGGTCGACGCGGTGCACAAGCTCAAATACGAGAATTACACCAGCAGCTTCTTCATCAGGGACATCATCAAGCCCGACCCCCCCAAGAACCTGCAGCTGAAGCCCTTGAAGAACAGCAGGCAGGTGGAGGTGAGCTGGGAGTACCCGGACACCTGGAGCACCCCCCACTCCTACTTCAGCCTGACGTTCTGTGTGCAGGTGCAGGGGAAGTCCAAGAGGGAGAAGAAGGACCGGGTGTTCACCGACAAGACCAGCGCCACCGTGATATGCCGCAAGAACGCGTCCATCAGCGTTCGCGCCCAGGACCGCTACTACAGCAGCTCCTGGTCCGAATGGGCCAGCGTGCCCTGCAGCGGTGGAGGGGGCGGGGGCTCCAGGAATCTGCCGGTGGCCACCCCCGACCCCGGGATGTTCCCGTGTCTGCATCACTCCCAGAACCTGCTGCGGGCCGTGAGCAATATGCTGCAGAAGGCCAGGCAGACGCTCGAGTTCTACCCCTGCACCTCCGAAGAGATCGACCATGAGGACATCACCAAGGACAAGACCAGCACCGTGGAGGCCTGCCTCCCCCTGGAGCTGACCAAAAACGAGAGCTGCCTGAACTCCAGGGAGACCAGCTTTATAACCAACGGCAGCTGCCTCGCCTCCAGGAAGACCTCGTTTATGATGGCCCTCTGCCTGTCCAGCATCTACGAGGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAACGCGAAGTTGCTCATGGACCCCAAGAGGCAGATCTTCCTGGACCAGAACATGCTCGCGGTGATCGACGAGCTGATGCAAGCCCTGAACTTCAACAGCGAGACCGTGCCCCAGAAGAGCAGCCTGGAAGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATCCTGCTGCACGCCTTCCGGATCCGGGCCGTGACCATCGACAGGGTGATGAGCTACCTCAACGCCTCCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 138 hIL12AB_038TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAGCAGCTCGT 3′UTR)GATCAGCTGGTTCTCCCTCGTCTTCCTGGCCTCCCCGCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGCTGGACTGGTATCCCGACGCCCCCGGCGAGATGGTGGTGCTGACGTGCGACACACCAGAAGAGGACGGGATCACATGGACCCTGGATCAGTCGTCCGAGGTGCTGGGGAGCGGCAAGACCCTCACCATCCAAGTGAAGGAGTTCGGGGACGCCGGCCAGTACACCTGCCACAAGGGCGGGGAGGTGCTCTCCCATAGCCTGCTCCTCCTGCACAAAAAGGAGGATGGCATCTGGAGCACCGACATCCTGAAGGACCAGAAGGAGCCCAAGAACAAGACATTTCTCAGGTGTGAGGCCAAGAACTATTCGGGCAGGTTTACCTGTTGGTGGCTCACCACCATCTCTACCGACCTGACGTTCTCCGTCAAGTCAAGCAGGGGGAGCTCGGACCCCCAGGGGGTGACATGTGGGGCCGCCACCCTGAGCGCGGAGCGTGTCCGCGGCGACAACAAGGAGTACGAGTATTCCGTGGAGTGCCAGGAGGACAGCGCCTGCCCCGCCGCCGAGGAGTCCCTGCCCATAGAGGTGATGGTGGACGCCGTCCACAAGTTGAAGTACGAAAATTATACCTCCTCGTTCTTCATTAGGGACATCATCAAGCCTGACCCCCCGAAGAACCTACAACTCAAGCCCCTCAAGAACTCCCGCCAGGTGGAGGTGTCCTGGGAGTACCCCGACACCTGGTCCACCCCGCACAGCTACTTCAGCCTGACCTTCTGCGTGCAGGTCCAGGGGAAGAGCAAGCGTGAAAAGAAAGACAGGGTGTTCACCGACAAGACGAGCGCCACCGTGATCTGCAGGAAAAACGCCTCCATCTCCGTGCGCGCCCAGGACAGGTACTACAGTAGCTCCTGGAGCGAATGGGCCAGCGTGCCGTGCAGCGGCGGGGGAGGAGGCGGCAGTCGCAACCTGCCCGTGGCCACCCCCGACCCCGGCATGTTCCCATGCCTGCACCACAGCCAGAACCTGCTGAGGGCAGTCAGCAATATGCTGCAGAAGGCCAGGCAGACCCTGGAGTTTTATCCCTGCACCAGCGAGGAGATCGACCACGAGGACATCACCAAGGACAAGACCTCCACCGTCGAGGCCTGCCTGCCACTGGAGCTGACCAAAAACGAGAGCTGCCTGAACTCCAGGGAGACCTCCTTCATCACCAACGGGAGCTGCCTGGCCAGCCGGAAGACCAGCTTCATGATGGCGCTGTGCCTCAGCAGCATCTACGAGGATCTCAAGATGTACCAGGTGGAGTTCAAGACCATGAACGCGAAGCTGCTGATGGACCCCAAGCGGCAGATCTTCCTGGACCAGAACATGCTGGCCGTGATTGACGAGCTCATGCAGGCCCTGAACTTCAATAGCGAGACCGTCCCCCAAAAGAGCAGCCTGGAGGAACCCGACTTCTACAAAACGAAGATCAAGCTCTGCATCCTGCTGCACGCCTTCCGGATCCGGGCCGTGACCATCGATCGTGTGATGAGCTACCTGAACGCCTCGTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 139 hIL12AB_039TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCACCAGCAGCTCGT 3′UTR)CATCTCCTGGTTTAGCCTGGTGTTTCTGGCCTCCCCCCTGGTCGCCATCTGGGAGCTGAAGAAAGACGTGTACGTGGTGGAGCTGGACTGGTACCCGGACGCTCCCGGGGAGATGGTGGTGCTGACCTGCGACACCCCCGAGGAGGACGGCATCACCTGGACCCTGGACCAGAGCTCCGAGGTGCTGGGGAGCGGCAAGACCCTGACCATTCAGGTGAAAGAGTTCGGCGACGCCGGCCAATATACCTGCCACAAGGGGGGGGAGGTCCTGTCGCATTCCCTGCTGCTGCTTCACAAAAAGGAGGATGGCATCTGGAGCACCGACATCCTGAAGGACCAGAAAGAACCCAAGAACAAGACGTTCCTGCGCTGCGAGGCCAAGAACTACAGCGGCCGGTTCACCTGTTGGTGGCTGACCACCATCTCCACCGACCTGACTTTCTCGGTGAAGAGCAGCCGCGGGAGCAGCGACCCCCAGGGAGTGACCTGCGGCGCCGCCACCCTGAGCGCCGAAAGGGTGAGGGGCGACAATAAAGAGTACGAGTATTCCGTGGAGTGCCAGGAGGACAGCGCCTGTCCCGCCGCCGAGGAGTCCCTGCCTATCGAGGTGATGGTCGACGCGGTGCACAAGCTCAAGTACGAAAACTACACCAGCAGCTTTTTCATCAGGGATATCATCAAACCAGACCCCCCCAAGAACCTGCAGCTGAAGCCCCTGAAAAACAGCAGGCAGGTGGAAGTGAGCTGGGAATACCCCGATACCTGGTCCACCCCCCACAGCTACTTCAGCCTGACCTTCTGCGTGCAGGTGCAGGGGAAGTCCAAGCGGGAGAAGAAAGATCGGGTGTTCACGGACAAGACCAGCGCCACCGTGATTTGCAGGAAAAACGCCAGCATCTCCGTGAGGGCTCAGGACAGGTACTACAGCTCCAGCTGGAGCGAGTGGGCCTCCGTGCCTTGCAGCGGGGGAGGAGGCGGCGGCAGCAGGAATCTGCCCGTCGCAACCCCCGACCCCGGCATGTTCCCCTGCCTGCACCACAGCCAGAATCTGCTGCGAGCCGTGAGCAACATGCTCCAGAAGGCCCGGCAGACGCTGGAGTTCTACCCCTGCACCTCCGAGGAGATCGACCACGAGGACATCACCAAGGATAAGACGAGCACCGTCGAGGCCTGTCTCCCCCTGGAGCTCACCAAGAACGAGTCCTGCCTGAATAGCAGGGAGACGTCCTTCATAACCAACGGCAGCTGTCTGGCGTCCAGGAAGACCAGCTTCATGATGGCCCTCTGCCTGAGCTCCATCTACGAGGACCTCAAGATGTACCAGGTCGAGTTCAAGACCATGAACGCAAAACTGCTCATGGATCCAAAGAGGCAGATCTTTCTGGACCAGAACATGCTGGCCGTGATCGATGAACTCATGCAGGCCCTGAATTTCAATTCCGAGACCGTGCCCCAGAAGAGCTCCCTGGAGGAACCCGACTTCTACAAAACAAAGATCAAGCTGTGTATCCTCCTGCACGCCTTCCGGATCAGGGCCGTCACCATTGACCGGGTGATGTCCTACCTGAACGCCAGCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 140 hIL12AB_040TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGA (5′UTR ORFGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGTGCCATCAGCAGCTGGT 3′UTR)GATCAGCTGGTTCAGCCTCGTGTTCCTCGCCAGCCCCCTCGTGGCCATCTGGGAGCTGAAAAAGGACGTGTACGTGGTGGAGCTGGACTGGTATCCCGACGCCCCGGGCGAGATGGTGGTGCTGACCTGCGACACCCCCGAGGAGGACGGCATTACCTGGACACTGGACCAGAGCAGCGAGGTCCTGGGCAGCGGGAAGACCCTGACAATTCAGGTGAAGGAGTTCGGCGACGCCGGACAGTACACGTGCCACAAGGGGGGGGAGGTGCTGTCCCACAGCCTCCTCCTGCTGCACAAGAAGGAGGATGGCATCTGGAGCACCGACATCCTGAAGGATCAGAAGGAGCCCAAGAACAAGACCTTTCTGAGATGCGAGGCCAAGAATTACAGCGGCCGTTTCACCTGCTGGTGGCTCACCACCATCAGCACCGACCTGACCTTCAGCGTGAAATCCTCCAGGGGCTCCTCCGACCCGCAGGGAGTGACCTGCGGCGCCGCCACACTGAGCGCCGAGCGGGTCAGAGGGGACAACAAGGAGTACGAGTACAGCGTTGAGTGCCAGGAGGACAGCGCCTGTCCCGCGGCCGAGGAATCCCTGCCCATCGAGGTGATGGTGGACGCAGTGCACAAGCTGAAGTACGAGAACTATACCTCGAGCTTCTTCATCCGGGATATCATTAAGCCCGATCCCCCGAAGAACCTGCAGCTCAAACCCCTGAAGAACAGCAGGCAGGTGGAGGTCTCCTGGGAGTACCCCGACACATGGTCCACCCCCCATTCCTATTTCTCCCTGACCTTTTGCGTGCAGGTGCAGGGCAAGAGCAAGAGGGAGAAAAAGGACAGGGTGTTCACCGACAAGACCTCCGCCACCGTGATCTGCCGTAAGAACGCTAGCATCAGCGTCAGGGCCCAGGACAGGTACTATAGCAGCTCCTGGTCCGAGTGGGCCAGCGTCCCGTGCAGCGGCGGGGGCGGTGGAGGCTCCCGGAACCTCCCCGTGGCCACCCCGGACCCCGGGATGTTTCCCTGCCTGCATCACAGCCAGAACCTGCTGAGGGCCGTGTCCAACATGCTGCAGAAGGCCAGGCAGACACTCGAGTTTTACCCCTGCACCAGCGAGGAGATCGACCACGAAGACATCACCAAGGACAAGACCTCCACCGTGGAGGCATGCCTGCCCCTGGAGCTGACCAAAAACGAAAGCTGTCTGAACTCCAGGGAGACCTCCTTTATCACGAACGGCTCATGCCTGGCCTCCAGAAAGACCAGCTTCATGATGGCCCTGTGCCTGAGCTCCATCTACGAGGACTTGAAAATGTACCAGGTCGAGTTCAAGACCATGAACGCCAAGCTGCTCATGGACCCCAAAAGGCAGATCTTTCTGGACCAGAATATGCTGGCCGTGATCGACGAGCTCATGCAAGCCCTGAATTTCAACAGCGAGACCGTGCCCCAGAAGTCCTCCCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATACTCCTGCACGCGTTTAGGATCAGGGCGGTGACCATCGATAGGGTGATGAGCTACCTGAATGCCTCCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCCAAACACCATTGTCACACTCCAGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC 141 hIL12AB_001G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGUC (mRNA withACCAGCAGCUGGUCAUUAGCUGGUUUAGCCUUGUGUUCCUGGCCUCCCCCCUUGUC T100 tail)GCUAUUUGGGAGCUCAAGAAGGACGUGUACGUGGUGGAGUUGGAUUGGUACCCAGACGCGCCCGGAGAGAUGGUAGUUCUGACCUGUGAUACCCCAGAGGAGGACGGCAUCACCUGGACGCUGGACCAAAGCAGCGAGGUUUUGGGCUCAGGGAAAACGCUGACCAUCCAGGUGAAGGAAUUCGGCGACGCCGGGCAGUACACCUGCCAUAAGGGAGGAGAGGUGCUGAGCCAUUCCCUUCUUCUGCUGCACAAGAAAGAGGACGGCAUCUGGUCUACCGACAUCCUGAAAGACCAGAAGGAGCCCAAGAACAAAACCUUCCUGAGGUGCGAGGCCAAGAACUACUCCGGCAGGUUCACUUGUUGGUGGCUGACCACCAUCAGUACAGACCUGACUUUUAGUGUAAAAAGCUCCAGAGGCUCGUCCGAUCCCCAAGGGGUGACCUGCGGCGCAGCCACUCUGAGCGCUGAGCGCGUGCGCGGUGACAAUAAAGAGUACGAGUACAGCGUUGAGUGUCAAGAAGAUAGCGCUUGCCCUGCCGCCGAGGAGAGCCUGCCUAUCGAGGUGAUGGUUGACGCAGUGCACAAGCUUAAGUACGAGAAUUACACCAGCUCAUUCUUCAUUAGAGAUAUAAUCAAGCCUGACCCACCCAAGAACCUGCAGCUGAAGCCACUGAAAAACUCACGGCAGGUCGAAGUGAGCUGGGAGUACCCCGACACCUGGAGCACUCCUCAUUCCUAUUUCUCUCUUACAUUCUGCGUCCAGGUGCAGGGCAAGAGCAAGCGGGAAAAGAAGGAUCGAGUCUUCACCGACAAAACAAGCGCGACCGUGAUUUGCAGGAAGAACGCCAGCAUCUCCGUCAGAGCCCAGGAUAGAUACUAUAGUAGCAGCUGGAGCGAGUGGGCAAGCGUGCCCUGUUCCGGCGGCGGGGGCGGGGGCAGCCGAAACUUGCCUGUCGCUACCCCGGACCCUGGAAUGUUUCCGUGUCUGCACCACAGCCAGAACCUGCUGAGAGCCGUGUCGAAUAUGCUCCAGAAGGCCCGGCAGACCCUUGAGUUCUACCCCUGUACCAGCGAAGAGAUCGAUCAUGAAGAUAUCACGAAAGAUAAAACAUCCACCGUCGAGGCUUGUCUCCCGCUGGAGCUGACCAAGAACGAGAGCUGUCUGAAUAGCCGGGAGACGUCUUUCAUCACGAAUGGUAGCUGUCUGGCCAGCAGGAAAACUUCCUUCAUGAUGGCUCUCUGCCUGAGCUCUAUCUAUGAAGAUCUGAAGAUGUAUCAGGUGGAGUUUAAAACAAUGAACGCCAAACUCCUGAUGGACCCAAAAAGGCAAAUCUUUCUGGACCAGAAUAUGCUGGCCGUGAUAGACGAGCUGAUGCAGGCACUGAACUUCAACAGCGAGACGGUGCCACAGAAAUCCAGCCUGGAGGAGCCUGACUUUUACAAAACUAAGAUCAAGCUGUGUAUCCUGCUGCACGCCUUUAGAAUCCGUGCCGUGACUAUCGACAGGGUGAUGUCAUACCUCAACGCUUCAUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 142 hIL12AB_002G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAGCAGCUGGUGAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCAGCCCCCUGGUG T100 tail)GCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGUUGGAUUGGUACCCCGACGCCCCCGGCGAGAUGGUGGUGCUGACCUGCGACACCCCCGAGGAGGACGGCAUCACCUGGACCCUGGACCAGAGCAGCGAGGUGCUGGGCAGCGGCAAGACCCUGACCAUCCAGGUGAAGGAGUUCGGCGACGCCGGCCAGUACACCUGCCACAAGGGCGGCGAGGUGCUGAGCCACAGCCUGCUGCUGCUGCACAAGAAGGAGGACGGCAUCUGGAGCACCGACAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGAGAUGCGAGGCCAAGAACUACAGCGGCAGAUUCACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUGACCUUCAGCGUGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGUGACCUGCGGCGCCGCCACCCUGAGCGCCGAGAGAGUGAGAGGCGACAACAAGGAGUACGAGUACAGCGUGGAGUGCCAGGAAGAUAGCGCCUGCCCCGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUGCACAAGCUGAAGUACGAGAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAGCCCGACCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCCGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGAGCACCCCCCACAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGCAAGAGCAAGAGAGAGAAGAAAGAUAGAGUGUUCACCGACAAGACCAGCGCCACCGUGAUCUGCAGAAAGAACGCCAGCAUCAGCGUGAGAGCCCAAGAUAGAUACUACAGCAGCAGCUGGAGCGAGUGGGCCAGCGUGCCCUGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCUGCCCGUGGCCACCCCCGACCCCGGCAUGUUCCCCUGCCUGCACCACAGCCAGAACCUGCUGAGAGCCGUGAGCAACAUGCUGCAGAAGGCCCGGCAGACCCUGGAGUUCUACCCCUGCACCAGCGAGGAGAUCGACCACGAAGAUAUCACCAAAGAUAAGACCAGCACCGUGGAGGCCUGCCUGCCCCUGGAGCUGACCAAGAACGAGAGCUGCCUGAACAGCAGAGAGACCAGCUUCAUCACCAACGGCAGCUGCCUGGCCAGCAGAAAGACCAGCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGGCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAGACCGUGCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCAGAAUCAGAGCCGUGACCAUCGACAGAGUGAUGAGCUACCUGAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 143 hIL12AB_003G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGUC (mRNA withACCAGCAGUUGGUCAUCUCUUGGUUUUCCCUGGUUUUUCUGGCAUCUCCCCUCGUG T100 tail)GCCAUCUGGGAACUGAAGAAAGACGUUUACGUUGUAGAAUUGGAUUGGUAUCCGGACGCUCCUGGAGAAAUGGUGGUCCUCACCUGUGACACCCCUGAAGAAGACGGAAUCACCUGGACCUUGGACCAGAGCAGUGAGGUCUUAGGCUCUGGCAAAACCCUGACCAUCCAAGUCAAAGAGUUUGGAGAUGCUGGCCAGUACACCUGUCACAAAGGAGGCGAGGUUCUAAGCCAUUCGCUCCUGCUGCUUCACAAAAAGGAAGAUGGAAUUUGGUCCACUGAUAUUUUAAAGGACCAGAAAGAACCCAAAAAUAAGACCUUUCUAAGAUGCGAGGCCAAGAAUUAUUCUGGACGUUUCACCUGCUGGUGGCUGACGACAAUCAGUACUGAUUUGACAUUCAGUGUCAAAAGCAGCAGAGGCUCUUCUGACCCCCAAGGGGUGACGUGCGGAGCUGCUACACUCUCUGCAGAGAGAGUCAGAGGUGACAACAAGGAGUAUGAGUACUCAGUGGAGUGCCAGGAAGAUAGUGCCUGCCCAGCUGCUGAGGAGAGUCUGCCCAUUGAGGUCAUGGUGGAUGCCGUUCACAAGCUCAAGUAUGAAAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAACCUGACCCACCCAAGAACUUGCAGCUGAAGCCAUUAAAGAAUUCUCGGCAGGUGGAGGUCAGCUGGGAGUACCCUGACACCUGGAGUACUCCACAUUCCUACUUCUCCCUGACAUUCUGCGUUCAGGUCCAGGGCAAGAGCAAGAGAGAAAAGAAAGAUAGAGUCUUCACAGAUAAGACCUCAGCCACGGUCAUCUGCCGCAAAAAUGCCAGCAUUAGCGUGCGGGCCCAGGACCGCUACUAUAGCUCAUCUUGGAGCGAAUGGGCAUCUGUGCCCUGCAGUGGCGGAGGGGGCGGAGGGAGCAGAAACCUCCCCGUGGCCACUCCAGACCCAGGAAUGUUCCCAUGCCUUCACCACUCCCAAAACCUGCUGAGGGCCGUCAGCAACAUGCUCCAGAAGGCCCGGCAAACUUUAGAAUUUUACCCUUGCACUUCUGAAGAGAUUGAUCAUGAAGAUAUCACAAAAGAUAAAACCAGCACAGUGGAGGCCUGUUUACCAUUGGAAUUAACCAAGAAUGAGAGUUGCCUAAAUUCCAGAGAGACCUCUUUCAUAACUAAUGGGAGUUGCCUGGCCUCCAGAAAGACCUCUUUUAUGAUGGCCCUGUGCCUUAGUAGUAUUUAUGAAGAUUUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAAUGCAAAGCUUCUGAUGGAUCCUAAGAGGCAGAUCUUUUUAGAUCAAAACAUGCUGGCAGUUAUUGAUGAGCUGAUGCAGGCCCUGAAUUUCAACAGUGAGACGGUGCCACAAAAAUCCUCCCUUGAAGAACCAGAUUUCUACAAGACCAAGAUCAAGCUCUGCAUACUUCUUCAUGCUUUCAGAAUUCGGGCAGUGACUAUUGAUAGAGUGAUGAGCUAUCUGAAUGCUUCCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 144 hIL12AB_004G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGGGCU (mRNA withGCCACCAGCAGCUGGUCAUCAGCUGGUUCUCCCUGGUCUUCCUGGCCAGCCCCCUG T100 tail)GUGGCCAUCUGGGAGCUGAAGAAAGACGUCUACGUAGUAGAGUUGGAUUGGUACCCAGACGCACCUGGAGAAAUGGUGGUUCUCACCUGUGACACGCCAGAAGAAGACGGUAUCACCUGGACGCUGGACCAGAGCUCAGAAGUUCUUGGCAGUGGAAAAACGCUGACCAUACAAGUAAAAGAAUUUGGGGAUGCUGGCCAGUACACCUGCCACAAAGGAGGAGAAGUUCUCAGCCACAGCCUGCUGCUGCUGCACAAGAAAGAAGAUGGCAUCUGGAGCACAGAUAUUUUAAAAGACCAGAAGGAGCCCAAGAACAAAACCUUCCUUCGAUGUGAGGCCAAGAACUACAGUGGCCGCUUCACCUGCUGGUGGCUCACCACCAUCAGCACAGACCUCACCUUCUCGGUGAAGAGCAGCCGUGGCAGCUCAGACCCCCAAGGAGUCACCUGUGGGGCGGCCACGCUGUCGGCAGAAAGAGUUCGAGGUGACAACAAGGAAUAUGAAUACUCGGUGGAAUGUCAAGAAGAUUCGGCCUGCCCGGCGGCAGAAGAAAGUCUUCCCAUAGAAGUCAUGGUGGAUGCUGUUCACAAAUUAAAAUAUGAAAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAGCCAGACCCGCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCCGGCAGGUGGAAGUUUCCUGGGAGUACCCAGAUACGUGGAGCACGCCGCACAGCUACUUCAGCCUCACCUUCUGUGUACAAGUACAAGGCAAGAGCAAGAGAGAGAAGAAAGAUCGUGUCUUCACAGAUAAAACCUCGGCGACGGUCAUCUGCAGGAAGAAUGCCUCCAUCUCGGUUCGAGCCCAGGACCGCUACUACAGCAGCAGCUGGAGUGAGUGGGCCUCGGUGCCCUGCAGUGGUGGCGGCGGCGGCGGCAGCAGAAACCUUCCUGUGGCCACGCCGGACCCUGGCAUGUUCCCGUGCCUGCACCACAGCCAAAAUUUACUUCGAGCUGUUUCUAACAUGCUGCAGAAAGCACGGCAAACUUUAGAAUUCUACCCCUGCACCUCAGAAGAAAUAGACCAUGAAGAUAUCACCAAAGAUAAAACCAGCACUGUAGAGGCCUGCCUGCCCCUGGAGCUCACCAAGAAUGAAUCCUGCCUCAACAGCAGAGAGACCAGCUUCAUCACCAAUGGCAGCUGCCUGGCCAGCAGGAAAACCAGCUUCAUGAUGGCGCUCUGCCUGAGCAGCAUCUAUGAAGAUUUGAAGAUGUACCAAGUAGAAUUUAAAACCAUGAAUGCCAAGCUGCUCAUGGACCCCAAGCGGCAGAUAUUUUUGGAUCAAAACAUGCUGGCUGUCAUUGAUGAGCUCAUGCAAGCAUUAAACUUCAACUCAGAGACGGUGCCCCAGAAGAGCAGCCUGGAGGAGCCAGAUUUCUACAAAACCAAGAUCAAGCUCUGCAUCUUAUUACAUGCCUUCCGCAUCCGGGCGGUCACCAUUGACCGUGUCAUGUCCUACUUAAAUGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 145 hIL12AB_005G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAGCAGCUGGUCAUCAGCUGGUUCUCCCUGGUCUUCCUGGCCAGCCCCCUGGUG T100 tail)GCCAUCUGGGAGCUGAAGAAAGACGUCUACGUAGUAGAGUUGGAUUGGUACCCAGACGCACCUGGAGAAAUGGUGGUUCUCACCUGUGACACGCCAGAAGAAGACGGUAUCACCUGGACGCUGGACCAGAGCUCAGAAGUUCUUGGCAGUGGAAAAACGCUGACCAUACAAGUAAAAGAAUUUGGGGAUGCUGGCCAGUACACCUGCCACAAAGGAGGAGAAGUUCUCAGCCACAGCCUGCUGCUGCUGCACAAGAAAGAAGAUGGCAUCUGGAGCACAGAUAUUUUAAAAGACCAGAAGGAGCCCAAGAACAAAACCUUCCUUCGAUGUGAGGCCAAGAACUACAGUGGCCGCUUCACCUGCUGGUGGCUCACCACCAUCAGCACAGACCUCACCUUCUCGGUGAAGAGCAGCCGUGGCAGCUCAGACCCCCAAGGAGUCACCUGUGGGGCGGCCACGCUGUCGGCAGAAAGAGUUCGAGGUGACAACAAGGAAUAUGAAUACUCGGUGGAAUGUCAAGAAGAUUCGGCCUGCCCGGCGGCAGAAGAAAGUCUUCCCAUAGAAGUCAUGGUGGAUGCUGUUCACAAAUUAAAAUAUGAAAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAGCCAGACCCGCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCCGGCAGGUGGAAGUUUCCUGGGAGUACCCAGAUACGUGGAGCACGCCGCACAGCUACUUCAGCCUCACCUUCUGUGUACAAGUACAAGGCAAGAGCAAGAGAGAGAAGAAAGAUCGUGUCUUCACAGAUAAAACCUCGGCGACGGUCAUCUGCAGGAAGAAUGCCUCCAUCUCGGUUCGAGCCCAGGACCGCUACUACAGCAGCAGCUGGAGUGAGUGGGCCUCGGUGCCCUGCAGUGGUGGCGGCGGCGGCGGCAGCAGAAACCUUCCUGUGGCCACGCCGGACCCUGGCAUGUUCCCGUGCCUGCACCACAGCCAAAAUUUACUUCGAGCUGUUUCUAACAUGCUGCAGAAAGCACGGCAAACUUUAGAAUUCUACCCCUGCACCUCAGAAGAAAUAGACCAUGAAGAUAUCACCAAAGAUAAAACCAGCACUGUAGAGGCCUGCCUGCCCCUGGAGCUCACCAAGAAUGAAUCCUGCCUCAACAGCAGAGAGACCAGCUUCAUCACCAAUGGCAGCUGCCUGGCCAGCAGGAAAACCAGCUUCAUGAUGGCGCUCUGCCUGAGCAGCAUCUAUGAAGAUUUGAAGAUGUACCAAGUAGAAUUUAAAACCAUGAAUGCCAAGCUGCUCAUGGACCCCAAGCGGCAGAUAUUUUUGGAUCAAAACAUGCUGGCUGUCAUUGAUGAGCUCAUGCAAGCAUUAAACUUCAACUCAGAGACGGUGCCCCAGAAGAGCAGCCUGGAGGAGCCAGAUUUCUACAAAACCAAGAUCAAGCUCUGCAUCUUAUUACAUGCCUUCCGCAUCCGGGCGGUCACCAUUGACCGUGUCAUGUCCUACUUAAAUGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 146 hIL12AB_006G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAGCAGCUGGUGAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCAGCCCCCUGGUG T100 tail)GCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGUUGGAUUGGUACCCCGACGCCCCCGGCGAGAUGGUGGUGCUGACCUGUGACACCCCCGAGGAGGACGGCAUCACCUGGACCCUGGACCAGAGCAGCGAGGUGCUGGGCAGCGGCAAGACCCUGACCAUCCAGGUGAAGGAGUUCGGGGACGCCGGCCAGUACACCUGCCACAAGGGCGGCGAGGUGCUGAGCCACAGCCUGCUGCUGCUGCACAAGAAGGAGGACGGCAUCUGGAGCACAGAUAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGAGAUGCGAGGCCAAGAACUACAGCGGCAGAUUCACCUGCUGGUGGCUGACCACCAUCAGCACAGAUUUGACCUUCAGCGUGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGUGACCUGCGGCGCCGCCACCCUGAGCGCCGAGAGAGUGAGAGGUGACAACAAGGAGUACGAGUACAGCGUGGAGUGCCAGGAAGAUAGCGCCUGCCCCGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUGCACAAGCUGAAGUACGAGAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAGCCCGACCCGCCGAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCCGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGAGCACCCCCCACAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGCAAGAGCAAGAGAGAGAAGAAAGAUAGAGUGUUCACAGAUAAGACCAGCGCCACCGUGAUCUGCAGAAAGAACGCCAGCAUCAGCGUGAGAGCCCAAGAUAGAUACUACAGCAGCAGCUGGAGCGAGUGGGCCAGCGUGCCCUGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCUGCCCGUGGCCACCCCCGACCCCGGCAUGUUCCCCUGCCUGCACCACAGCCAGAACCUGCUGAGAGCCGUGAGCAACAUGCUGCAGAAGGCCCGGCAGACCCUGGAGUUCUACCCCUGCACCAGCGAGGAGAUCGACCACGAAGAUAUCACCAAAGAUAAGACCAGCACCGUGGAGGCCUGCCUGCCCCUGGAGCUGACCAAGAAUGAAAGCUGCCUGAACAGCAGAGAGACCAGCUUCAUCACCAACGGCAGCUGCCUGGCCAGCAGAAAGACCAGCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGGCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAGACCGUGCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCAGAAUCAGAGCCGUGACCAUCGACAGAGUGAUGAGCUACCUGAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 147 hIL12AB_007G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAGCAGCUUGUCAUCUCCUGGUUCUCUCUUGUCUUCCUUGCUUCUCCUCUUGUG T100 tail)GCCAUCUGGGAGCUGAAGAAGGACGUUUACGUAGUGGAGUUGGAUUGGUACCCUGACGCACCUGGAGAAAUGGUGGUUCUCACCUGUGACACUCCUGAGGAGGACGGUAUCACCUGGACGUUGGACCAGUCUUCUGAGGUUCUUGGCAGUGGAAAAACUCUUACUAUUCAGGUGAAGGAGUUUGGAGAUGCUGGCCAGUACACCUGCCACAAGGGUGGUGAAGUUCUCAGCCACAGUUUACUUCUUCUUCACAAGAAGGAGGAUGGCAUCUGGUCUACUGACAUUUUAAAAGACCAGAAGGAGCCCAAGAACAAAACAUUCCUUCGUUGUGAAGCCAAGAACUACAGUGGUCGUUUCACCUGCUGGUGGCUUACUACUAUUUCUACUGACCUUACUUUCUCUGUGAAGUCUUCUCGUGGCUCUUCUGACCCUCAGGGUGUCACCUGUGGGGCUGCUACUCUUUCUGCUGAGCGUGUGCGUGGUGACAACAAGGAGUAUGAAUACUCGGUGGAGUGCCAGGAAGAUUCUGCCUGCCCUGCUGCUGAGGAGUCUCUUCCUAUUGAGGUGAUGGUGGAUGCUGUGCACAAGUUAAAAUAUGAAAACUACACUUCUUCUUUCUUCAUUCGUGACAUUAUAAAACCUGACCCUCCCAAGAACCUUCAGUUAAAACCUUUAAAAAACUCUCGUCAGGUGGAGGUGUCCUGGGAGUACCCUGACACGUGGUCUACUCCUCACUCCUACUUCUCUCUUACUUUCUGUGUCCAGGUGCAGGGCAAGUCCAAGCGUGAGAAGAAGGACCGUGUCUUCACUGACAAAACAUCUGCUACUGUCAUCUGCAGGAAGAAUGCAUCCAUCUCUGUGCGUGCUCAGGACCGUUACUACAGCUCUUCCUGGUCUGAGUGGGCUUCUGUGCCCUGCUCUGGCGGCGGCGGCGGCGGCAGCAGAAAUCUUCCUGUGGCUACUCCUGACCCUGGCAUGUUCCCCUGCCUUCACCACUCGCAGAACCUUCUUCGUGCUGUGAGCAACAUGCUUCAGAAGGCUCGUCAAACUUUAGAAUUCUACCCCUGCACUUCUGAGGAGAUUGAGCAUGAAGAUAUCACCAAAGAUAAAACAUCUACUGUGGAGGCCUGCCUUCCUUUAGAGCUGACCAAGAAUGAAUCCUGCUUAAAUUCUCGUGAGACGUCUUUCAUCACCAAUGGCAGCUGCCUUGCCUCGCGCAAAACAUCUUUCAUGAUGGCUCUUUGCCUUUCUUCCAUCUAUGAAGAUUUAAAAAUGUACCAGGUGGAGUUCAAGACCAUGAAUGCAAAGCUUCUCAUGGACCCCAAGCGUCAGAUAUUUUUGGACCAGAACAUGCUUGCUGUCAUUGAUGAGCUCAUGCAGGCUUUAAACUUCAACUCUGAGAGGGUGCCUCAGAAGUCUUCUUUAGAAGAGCCUGAGUUCUACAAGACCAAGAUAAAACUUUGCAUUCUUCUUCAUGCUUUCCGCAUCCGUGCUGUGACUAUUGACCGUGUGAUGUCCUACUUAAAUGCUUCUUGAUAAUAGGCUGGAGCCUCGGUGGCCAAGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 148 hIL12AB_008G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGUC (mRNA withAUCAACAACUCGUGAUUAGCUGGUUCAGUCUCGUGUUCCUGGCCUCUCCGCUGGUG T100 tail)GCCAUCUGGGAGCUUAAGAAGGACGUGUACGUGGUGGAGCUCGAUUGGUACCCCGACGCACCUGGCGAGAUGGUGGUGCUAACCUGCGAUACCCCCGAGGAGGACGGGAUCACUUGGACCCUGGAUCAGAGUAGCGAAGUCCUGGGCUCUGGCAAAACACUCACAAUCCAGGUGAAGGAAUUCGGAGACGCUGGUCAGUACACUUGCCACAAGGGGGGUGAAGUGCUGUCUCACAGCCUGCUGUUACUGCACAAGAAGGAGGAUGGGAUCUGGUCAACCGACAUCCUGAAGGAUCAGAAGGAGCCUAAGAACAAGACCUUUCUGAGGUGUGAAGCUAAGAACUAUUCCGGAAGAUUCACUUGCUGGUGGUUGACCACAAUCAGCACUGACCUGACCUUUUCCGUGAAGUCCAGCAGAGGAAGCAGCGAUCCUCAGGGCGUAACGUGCGGCGCGGCUACCCUGUCAGCUGAGCGGGUUAGAGGCGACAACAAAGAGUAUGAGUACUCCGUGGAGUGUCAGGAAGAUAGCGCCUGCCCCGCAGCCGAGGAGAGUCUGCCCAUCGAGGUGAUGGUGGACGCUGUCCAUAAGUUAAAAUACGAAAAUUACACAAGUUCCUUUUUCAUCCGCGAUAUUAUCAAACCCGAUCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAAUAGCCGACAGGUGGAAGUCUCUUGGGAGUAUCCUGACACCUGGUCCACGCCUCACAGCUACUUUAGUCUGACUUUCUGUGUCCAGGUCCAGGGCAAGAGCAAGAGAGAGAAAAAGGAUAGAGUGUUUACUGACAAAACAUCUGCUACAGUCAUCUGCAGAAAGAACGCCAGUAUCUCAGUGAGGGCGCAAGAUAGAUACUACAGUAGUAGCUGGAGCGAAUGGGCUAGCGUGCCCUGUUCAGGGGGCGGCGGAGGGGGCUCCAGGAAUCUGCCCGUGGCCACCCCCGACCCUGGGAUGUUCCCUUGCCUCCAUCACUCACAGAACCUGCUCAGAGCAGUGAGCAACAUGCUCCAAAAGGCCCGCCAGACCCUGGAGUUUUACCCUUGUACUUCAGAAGAGAUCGAUCACGAAGAUAUAACAAAGGAUAAAACCAGCACCGUGGAGGCCUGUCUGCCUCUGGAACUCACAAAGAAUGAAAGCUGUCUGAAUUCCAGGGAAACCUCCUUCAUUACUAACGGAAGCUGUCUCGCAUCUCGCAAAACAUCAUUCAUGAUGGCCCUCUGCCUGUCUUCUAUCUAUGAAGAUCUCAAGAUGUAUCAGGUGGAGUUCAAAACAAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGGCAGAUCUUCCUGGACCAGAACAUGCUGGCAGUGAUCGAUGAGCUGAUGCAAGCCUUGAACUUCAACUCAGAGACGGUGCCGCAAAAGUCCUCGUUGGAGGAACCAGAUUUUUACAAAACCAAAAUCAAGCUGUGUAUCCUUCUUCACGCCUUUCGGAUCAGAGCCGUGACUAUCGACCGGGUGAUGUCAUACCUGAAUGCUUCCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 149 hIL12AB_009G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAGCAGCUGGUCAUCAGCUGGUUUAGCCUGGUCUUCCUGGCCAGCCCCCUGGUG T100 tail)GCCAUCUGGGAGCUGAAGAAAGACGUCUACGUAGUAGAGUUGGAUUGGUACCCAGACGCACCUGGAGAAAUGGUGGUUCUCACCUGCGACACGCCAGAAGAAGACGGUAUCACCUGGACGCUGGACCAGAGCAGCGAAGUACUGGGCAGUGGAAAAACGCUGACCAUACAAGUAAAAGAAUUUGGCGAUGCUGGCCAGUACACCUGCCACAAAGGAGGAGAAGUACUGAGCCACAGCCUGCUGCUGCUGCACAAGAAAGAAGAUGGCAUCUGGAGCACCGACAUUUUAAAAGACCAGAAGGAGCCCAAGAACAAAACCUUCCUUCGAUGUGAGGCGAAGAACUACAGUGGCCGCUUCACCUGCUGGUGGCUCACCACCAUCAGCACCGACCUCACCUUCUCGGUGAAGAGCAGCCGUGGUAGCUCAGACCCCCAAGGAGUCACCUGUGGGGCGGCCACGCUGUCGGCAGAAAGAGUUCGAGGCGACAACAAGGAAUAUGAAUACUCGGUGGAAUGUCAAGAAGAUUCGGCCUGCCCGGCGGCAGAAGAAAGUCUGCCCAUAGAAGUCAUGGUGGAUGCUGUUCACAAAUUAAAAUAUGAAAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAGCCAGACCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCCGGCAGGUGGAAGUUUCCUGGGAGUACCCAGAUACGUGGAGCACGCCGCACAGCUACUUCAGCCUCACCUUCUGUGUACAAGUACAAGGCAAGAGCAAGAGAGAGAAGAAAGAUCGUGUCUUCACCGACAAAACCUCGGCGACGGUCAUCUGCAGGAAGAAUGCAAGCAUCUCGGUUCGAGCCCAGGACCGCUACUACAGCAGCAGCUGGAGUGAGUGGGCCUCGGUGCCCUGCAGUGGUGGCGGCGGCGGCGGCAGCAGAAACCUUCCUGUGGCCACGCCGGACCCUGGCAUGUUUCCGUGCCUGCACCACAGCCAAAAUUUAUUACGAGCUGUUAGCAACAUGCUGCAGAAAGCACGGCAAACUUUAGAAUUCUACCCCUGCACCUCAGAAGAAAUAGACCAUGAAGAUAUCACCAAAGAUAAAACCAGCACUGUAGAGGCCUGCCUGCCCCUGGAGCUCACCAAGAACGAGAGCUGCCUCAAUAGCAGAGAGACCAGCUUCAUCACCAAUGGCAGCUGCCUGGCCAGCAGGAAAACCAGCUUCAUGAUGGCGCUCUGCCUGAGCAGCAUCUAUGAAGAUCUGAAGAUGUACCAAGUAGAAUUUAAAACCAUGAAUGCCAAGCUGCUCAUGGACCCCAAGCGGCAGAUAUUCCUCGACCAAAACAUGCUGGCUGUCAUUGAUGAGCUCAUGCAAGCAUUAAACUUCAACUCAGAGACGGUGCCCCAGAAGAGCAGCCUGGAGGAGCCAGAUUUCUACAAAACCAAGAUCAAGCUCUGCAUCUUAUUACAUGCCUUCCGCAUCCGGGCGGUCACCAUUGACCGUGUCAUGUCCUACUUAAAUGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 150 hIL12AB_010G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAGCAGCUUGUCAUCUCCUGGUUUUCUCUUGUCUUCCUCGCUUCUCCUCUUGUG T100 tail)GCCAUCUGGGAGCUGAAGAAAGACGUCUACGUAGUAGAGUUGGAUUGGUACCCGGACGCUCCUGGAGAAAUGGUGGUUCUCACCUGCGACACUCCUGAAGAAGACGGUAUCACCUGGACGCUGGACCAAAGCAGCGAAGUUUUAGGCUCUGGAAAAACGCUGACCAUACAAGUAAAAGAAUUUGGCGACGCUGGCCAGUACACGUGCCACAAAGGAGGAGAAGUUUUAAGCCACAGUUUACUUCUUCUUCACAAGAAAGAAGAUGGCAUCUGGAGUACAGAUAUUUUAAAAGACCAGAAGGAGCCUAAGAACAAAACCUUCCUCCGCUGUGAAGCUAAGAACUACAGUGGUCGUUUCACCUGCUGGUGGCUCACCACCAUCUCCACUGACCUCACCUUCUCUGUAAAAUCAAGCCGUGGUUCUUCUGACCCCCAAGGAGUCACCUGUGGGGCUGCCACGCUCAGCGCUGAAAGAGUUCGAGGCGACAACAAGGAAUAUGAAUAUUCUGUGGAAUGUCAAGAAGAUUCUGCCUGCCCGGCGGCAGAAGAAAGUCUUCCCAUAGAAGUCAUGGUGGACGCUGUUCACAAAUUAAAAUAUGAAAACUACACCAGCAGCUUCUUCAUUCGUGACAUCAUCAAACCAGACCCUCCUAAGAACCUUCAGUUAAAACCGCUGAAGAACAGCCGGCAGGUGGAAGUUUCCUGGGAGUACCCAGAUACGUGGAGUACGCCGCACUCCUACUUCAGUUUAACCUUCUGUGUACAAGUACAAGGAAAAUCAAAAAGAGAGAAGAAAGAUCGUGUCUUCACUGACAAAACAUCUGCCACGGUCAUCUGCCGUAAGAACGCUUCCAUCUCGGUUCGAGCCCAGGACCGCUACUACAGCAGCAGCUGGAGUGAGUGGGCAUCUGUUCCCUGCAGUGGUGGCGGCGGCGGCGGCAGCCGCAACCUUCCUGUGGCCACGCCGGACCCUGGCAUGUUCCCGUGCCUUCACCACUCGCAAAAUCUUCUUCGUGCUGUUUCUAACAUGCUGCAGAAGGCGCGGCAAACUUUAGAAUUCUACCCGUGCACUUCUGAAGAAAUAGACCAUGAAGAUAUCACCAAAGAUAAAACCAGCACGGUGGAGGCCUGCCUUCCUUUAGAACUUACUAAGAACGAAAGUUGCCUUAACAGCCGUGAGACCAGCUUCAUCACCAAUGGCAGCUGCCUUGCUAGCAGGAAGACCAGCUUCAUGAUGGCGCUGUGCCUUUCUUCCAUCUAUGAAGAUCUUAAGAUGUACCAAGUAGAAUUUAAAACCAUGAAUGCCAAAUUAUUAAUGGACCCCAAGCGGCAGAUAUUCCUCGACCAAAACAUGCUGGCUGUCAUUGAUGAGCUCAUGCAAGCAUUAAACUUCAACUCAGAAACUGUUCCCCAGAAGUCAUCUUUAGAAGAACCAGAUUUCUACAAAACAAAAAUAAAACUCUGCAUUCUUCUUCAUGCCUUCCGCAUCCGUGCUGUCACCAUUGACCGUGUCAUGUCCUACUUAAAUGCUUCUUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 151 hIL12AB_011G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAGCAGCUGGUGAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCAGCCCCCUGGUG T100 tail)GCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGUUGGAUUGGUACCCGGACGCGCCGGGGGAGAUGGUGGUGCUGACGUGCGACACGCCGGAGGAGGACGGGAUCACGUGGACGCUGGACCAGAGCAGCGAGGUGCUGGGGAGCGGGAAGACGCUGACGAUCCAGGUGAAGGAGUUCGGGGACGCGGGGCAGUACACGUGCCACAAGGGGGGGGAGGUGCUGAGCCACAGCCUGCUGCUGCUGCACAAGAAGGAGGACGGGAUCUGGAGCACAGAUAUCCUGAAGGACCAGAAGGAGCCGAAGAACAAGACGUUCCUGAGGUGCGAGGCGAAGAACUACAGCGGGAGGUUCACGUGCUGGUGGCUGACGACGAUCAGCACGGACCUGACGUUCAGCGUGAAGAGCAGCAGGGGGAGCAGCGACCCGCAGGGGGUGACGUGCGGGGCGGCGACGCUGAGCGCGGAGAGGGUGAGGGGUGACAACAAGGAGUACGAGUACAGCGUGGAGUGCCAGGAAGAUAGCGCGUGCCCGGCGGCGGAGGAGAGCCUGCCGAUCGAGGUGAUGGUGGACGCGGUGCACAAGCUGAAGUACGAGAACUACACGAGCAGCUUCUUCAUCAGAGAUAUCAUCAAGCCGGACCCGCCGAAGAACCUGCAGCUGAAGCCGCUGAAGAACAGCAGGCAGGUGGAGGUGAGCUGGGAGUACCCAGAUACGUGGAGCACGCCGCACAGCUACUUCAGCCUGACGUUCUGCGUGCAGGUGCAGGGGAAGAGCAAGAGGGAGAAGAAAGAUAGGGUGUUCACAGAUAAGACGAGCGCGACGGUGAUCUGCAGGAAGAACGCGAGCAUCAGCGUGAGGGCGCAAGAUAGGUACUACAGCAGCAGCUGGAGCGAGUGGGCGAGCGUGCCGUGCAGCGGGGGGGGGGGGGGGGGGAGCAGGAACCUGCCGGUGGCGACGCCGGACCCGGGGAUGUUCCCGUGCCUGCACCACAGCCAGAACCUGCUGAGGGCGGUGAGCAACAUGCUGCAGAAGGCGAGGCAGACGCUGGAGUUCUACCCGUGCACGAGCGAGGAGAUCGACCACGAAGAUAUCACGAAAGAUAAGACGAGCACGGUGGAGGCGUGCCUGCCGCUGGAGCUGACGAAGAACGAGAGCUGCCUGAACAGCAGGGAGACGAGCUUCAUCACGAACGGGAGCUGCCUGGCGAGCAGGAAGACGAGCUUCAUGAUGGCGCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACGAUGAACGCGAAGCUGCUGAUGGACCCGAAGAGGCAGAUCUUCCUGGACCAGAACAUGCUGGCGGUGAUCGACGAGCUGAUGCAGGCGCUGAACUUCAACAGCGAGACGGUGCCGCAGAAGAGCAGCCUGGAGGAGCCAGAUUUCUACAAGACGAAGAUCAAGCUGUGCAUCCUGCUGCACGCGUUCAGGAUCAGGGCGGUGACGAUCGACAGGGUGAUGAGCUACCUGAACGCGAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 152 hIL12AB_012G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withAUCAGCAGCUGGUGAUCAGCUGGUUCAGCCUCGUGUUUCUGGCCAGCCCCCUGGUG T100 tail)GCCAUUUGGGAACUCAAGAAGGACGUGUACGUUGUGGAACUCGACUGGUACCCUGACGCCCCAGGCGAAAUGGUGGUCUUAACCUGCGACACCCCUGAGGAGGACGGAAUCACCUGGACCUUGGACCAGAGCUCCGAGGUCCUCGGCAGUGGCAAGACCCUGACCAUACAGGUGAAAGAAUUUGGAGACGCAGGGCAAUACACAUGUCACAAGGGCGGGGAGGUUCUUUCUCACUCCCUUCUGCUUCUACAUAAAAAGGAAGACGGAAUUUGGUCUACCGACAUCCUCAAGGACCAAAAGGAGCCUAAGAAUAAAACCUUCUUACGCUGUGAAGCUAAAAACUACAGCGGCAGAUUCACUUGCUGGUGGCUCACCACCAUUUCUACCGACCUGACCUUCUCGGUGAAGUCUUCAAGGGGCUCUAGUGAUCCACAGGGAGUGACAUGCGGGGCCGCCACACUGAGCGCUGAACGGGUGAGGGGCGAUAACAAGGAGUAUGAAUACUCUGUCGAGUGUCAGGAGGAUUCAGCUUGUCCCGCAGCUGAAGAGUCACUCCCCAUAGAGGUUAUGGUCGAUGCUGUGCAUAAACUGAAGUACGAAAACUACACCAGCAGCUUCUUCAUUAGAGAUAUUAUAAAACCUGACCCCCCCAAGAACCUGCAACUUAAACCCCUGAAAAACUCUCGGCAGGUCGAAGUUAGCUGGGAGUACCCUGAUACUUGGUCCACCCCCCACUCGUACUUCUCACUGACUUUCUGUGUGCAGGUGCAGGGCAAGAGCAAGAGAGAGAAAAAAGAUCGUGUAUUCACAGAUAAGACCUCUGCCACCGUGAUCUGCAGAAAAAACGCUUCCAUCAGUGUCAGAGCCCAAGACCGGUACUAUAGUAGUAGCUGGAGCGAGUGGGCAAGUGUCCCCUGCUCUGGCGGCGGAGGGGGCGGCUCUCGAAACCUCCCCGUCGCUACCCCUGAUCCAGGAAUGUUCCCUUGCCUGCAUCACUCACAGAAUCUGCUGAGAGCGGUCAGCAACAUGCUGCAGAAAGCUAGGCAAACACUGGAGUUUUAUCCUUGUACCUCAGAGGAGAUCGACCACGAGGAUAUUACCAAAGAUAAGACCAGCACGGUGGAGGCCUGCUUGCCCCUGGAACUGACAAAGAAUGAAUCCUGCCUUAAUAGCCGUGAGACCUCUUUUAUAACAAACGGAUCCUGCCUGGCCAGCAGGAAGACCUCCUUCAUGAUGGCCCUCUGCCUGUCCUCAAUCUACGAAGACCUGAAGAUGUACCAGGUGGAAUUUAAAACUAUGAACGCCAAGCUGUUGAUGGACCCCAAGCGGCAGAUCUUUCUGGAUCAAAAUAUGCUGGCUGUGAUCGACGAACUGAUGCAGGCCCUCAACUUUAACAGCGAGACCGUGCCACAAAAGAGCAGUCUUGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUCCUUCAUGCCUUCAGGAUAAGAGCUGUCACCAUCGACAGAGUCAUGAGUUACCUGAAUGCAUCCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 153 hIL12AB_013G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAGCAGCUGGUCAUCUCCUGGUUCAGUCUUGUCUUCCUGGCCUCGCCGCUGGUG T100 tail)GCCAUCUGGGAGCUGAAGAAAGACGUUUACGUAGUAGAGUUGGAUUGGUACCCAGACGCACCUGGAGAAAUGGUGGUCCUCACCUGUGACACGCCAGAAGAAGACGGUAUCACCUGGACGCUGGACCAGAGCAGUGAAGUUCUUGGAAGUGGAAAAACGCUGACCAUACAAGUAAAAGAAUUUGGAGAUGCUGGCCAGUACACCUGCCACAAAGGAGGAGAAGUUCUCAGCCACAGUUUAUUAUUACUUCACAAGAAAGAAGAUGGCAUCUGGUCCACAGAUAUUUUAAAAGACCAGAAGGAGCCCAAAAAUAAAACAUUUCUUCGAUGUGAGGCCAAGAACUACAGUGGUCGUUUCACCUGCUGGUGGCUGACCACCAUCUCCACAGACCUCACCUUCAGUGUAAAAAGCAGCCGUGGUUCUUCUGACCCCCAAGGAGUCACCUGUGGGGCUGCCACGCUCUCUGCAGAAAGAGUUCGAGGUGACAACAAAGAAUAUGAGUACUCGGUGGAAUGUCAAGAAGAUUCGGCCUGCCCAGCUGCUGAGGAGAGUCUUCCCAUAGAAGUCAUGGUGGAUGCUGUUCACAAAUUAAAAUAUGAAAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAACCUGACCCGCCCAAGAACUUACAGCUGAAGCCGCUGAAAAACAGCCGGCAGGUAGAAGUUUCCUGGGAGUACCCAGAUACCUGGUCCACGCCGCACUCCUACUUCUCCCUCACCUUCUGUGUACAAGUACAAGGCAAGAGCAAGAGAGAGAAGAAAGAUCGUGUCUUCACAGAUAAAACAUCAGCCACGGUCAUCUGCAGGAAAAAUGCCAGCAUCUCGGUGCGGGCCCAGGACCGCUACUACAGCAGCAGCUGGAGUGAGUGGGCAUCUGUGCCCUGCAGUGGUGGUGGGGGUGGUGGCAGCAGAAACCUUCCUGUGGCCACUCCAGACCCUGGCAUGUUCCCGUGCCUUCACCACUCCCAAAAUUUACUUCGAGCUGUUUCUAACAUGCUGCAGAAAGCACGGCAAACUUUAGAAUUCUACCCGUGCACUUCUGAAGAAAUUGACCAUGAAGAUAUCACAAAAGAUAAAACCAGCACAGUGGAGGCCUGUCUUCCUUUAGAGCUGACCAAAAAUGAAUCCUGCCUCAACAGCAGAGAGACCAGCUUCAUCACCAAUGGCAGCUGCCUGGCCUCCAGGAAAACCAGCUUCAUGAUGGCGCUCUGCCUCAGCUCCAUCUAUGAAGAUUUGAAGAUGUACCAAGUAGAAUUUAAAACCAUGAAUGCCAAAUUAUUAAUGGACCCCAAGAGGCAGAUAUUUUUAGAUCAAAACAUGCUGGCAGUUAUUGAUGAGCUCAUGCAAGCAUUAAACUUCAACAGUGAGACGGUACCUCAAAAAAGCAGCCUUGAAGAGCCAGAUUUCUACAAAACCAAGAUCAAACUCUGCAUUUUACUUCAUGCCUUCCGCAUCCGGGCGGUCACCAUUGACCGUGUCAUGUCCUACUUAAAUGCCUCGUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 154 hIL12AB_014G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAGCAGCUUGUGAUUUCUUGGUUCUCUCUUGUGUUCCUUGCUUCUCCUCUUGUG T100 tail)GCUAUUUGGGAGUUAAAAAAGGACGUGUACGUGGUGGAGCUUGACUGGUACCCUGACGCACCUGGCGAGAUGGUGGUGCUUACUUGUGACACUCCUGAGGAGGACGGCAUUACUUGGACGCUUGACCAGUCUUCUGAGGUGCUUGGCUCUGGCAAAACACUUACUAUUCAGGUGAAGGAGUUCGGGGAUGCUGGCCAGUACACUUGCCACAAGGGCGGCGAGGUGCUUUCUCACUCUCUUCUUCUUCUUCACAAGAAGGAGGACGGCAUUUGGUCUACUGACAUUUUAAAAGACCAGAAGGAGCCCAAGAACAAAACAUUCCUUCGUUGCGAGGCCAAGAACUACUCUGGCCGUUUCACUUGCUGGUGGCUUACUACUAUUUCUACUGACCUUACUUUCUCUGUGAAGUCUUCUCGUGGCUCUUCUGACCCUCAGGGCGUGACUUGUGGGGCUGCUACUCUUUCUGCUGAGCGUGUGCGUGGUGACAACAAGGAGUACGAGUACUCUGUGGAGUGCCAGGAAGAUUCUGCUUGCCCUGCUGCUGAGGAGUCUCUUCCUAUUGAGGUGAUGGUGGAUGCUGUGCACAAGUUAAAAUACGAGAACUACACUUCUUCUUUCUUCAUUCGUGACAUUAUUAAGCCUGACCCUCCCAAGAACCUUCAGUUAAAACCUUUAAAAAACUCUCGUCAGGUGGAGGUGUCUUGGGAGUACCCUGACACUUGGUCUACUCCUCACUCUUACUUCUCUCUUACUUUCUGCGUGCAGGUGCAGGGCAAGUCUAAGCGUGAGAAGAAGGACCGUGUGUUCACUGACAAAACAUCUGCUACUGUGAUUUGCAGGAAGAAUGCAUCUAUUUCUGUGCGUGCUCAGGACCGUUACUACUCUUCUUCUUGGUCUGAGUGGGCUUCUGUGCCUUGCUCUGGCGGCGGCGGCGGCGGCUCCAGAAAUCUUCCUGUGGCUACUCCUGACCCUGGCAUGUUCCCUUGCCUUCACCACUCUCAGAACCUUCUUCGUGCUGUGAGCAACAUGCUUCAGAAGGCUCGUCAAACUCUUGAGUUCUACCCUUGCACUUCUGAGGAGAUUGACCACGAAGAUAUCACCAAAGAUAAAACAUCUACUGUGGAGGCUUGCCUUCCUCUUGAGCUUACCAAGAAUGAAUCUUGCUUAAAUUCUCGUGAGACGUCUUUCAUCACCAACGGCUCUUGCCUUGCCUCGCGCAAAACAUCUUUCAUGAUGGCUCUUUGCCUUUCUUCUAUUUACGAAGAUUUAAAAAUGUACCAGGUGGAGUUCAAAACAAUGAAUGCAAAGCUUCUUAUGGACCCCAAGCGUCAGAUUUUCCUUGACCAGAACAUGCUUGCUGUGAUUGACGAGCUUAUGCAGGCUUUAAAUUUCAACUCUGAGACGGUGCCUCAGAAGUCUUCUCUUGAGGAGCCUGACUUCUACAAGACCAAGAUUAAGCUUUGCAUUCUUCUUCAUGCUUUCCGUAUUCGUGCUGUGACUAUUGACCGUGUGAUGUCUUACUUAAAUGCUUCUUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 155 hIL12AB_015G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGUC (mRNA withACCAGCAGCUGGUGAUCAGCUGGUUUAGCCUGGUGUUUCUGGCCAGCCCCCUGGUG T100 tail)GCCAUCUGGGAACUGAAGAAAGACGUGUACGUGGUAGAACUGGAUUGGUAUCCGGACGCUCCCGGCGAAAUGGUGGUGCUGACCUGUGACACCCCCGAAGAAGACGGAAUCACCUGGACCCUGGACCAGAGCAGCGAGGUGCUGGGCAGCGGCAAAACCCUGACCAUCCAAGUGAAAGAGUUUGGCGAUGCCGGCCAGUACACCUGUCACAAAGGCGGCGAGGUGCUAAGCCAUUCGCUGCUGCUGCUGCACAAAAAGGAAGAUGGCAUCUGGAGCACCGAUAUCCUGAAGGACCAGAAAGAACCCAAAAAUAAGACCUUUCUAAGAUGCGAGGCCAAGAAUUAUAGCGGCCGUUUCACCUGCUGGUGGCUGACGACCAUCAGCACCGAUCUGACCUUCAGCGUGAAAAGCAGCAGAGGCAGCAGCGACCCCCAAGGCGUGACGUGCGGCGCCGCCACCCUGAGCGCCGAGAGAGUGAGAGGCGACAACAAGGAGUAUGAGUACAGCGUGGAGUGCCAGGAAGAUAGCGCCUGCCCCGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGAUGCCGUGCACAAGCUGAAGUAUGAAAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAACCCGAGCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAAUAGCCGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGAGCACCCCCCAUAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGCAAGAGCAAGAGAGAAAAGAAAGAUAGAGUGUUCACAGAUAAGACCAGCGCCACGGUGAUCUGCAGAAAAAAUGCCAGCAUCAGCGUGAGAGCCCAAGAUAGAUACUAUAGCAGCAGCUGGAGCGAAUGGGCCAGCGUGCCCUGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCUGCCCGUGGCCACCCCCGACCCCGGCAUGUUCCCCUGCCUGCACCACAGCCAAAACCUGCUGAGAGCCGUGAGCAACAUGCUGCAGAAGGCCCGGCAGACCCUGGAAUUUUACCCCUGCACCAGCGAAGAGAUCGAUCAUGAAGAUAUCACCAAAGAUAAAACCAGCACCGUGGAGGCCUGUCUGCCCCUGGAACUGACCAAGAAUGAGAGCUGCCUAAAUAGCAGAGAGACCAGCUUCAUAACCAAUGGCAGCUGCCUGGCCAGCAGAAAGACCAGCUUUAUGAUGGCCCUGUGCCUGAGCAGCAUCUAUGAAGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAAUGCCAAGCUGCUGAUGGAUCCCAAGCGGCAGAUCUUUCUGGAUCAAAACAUGCUGGCCGUGAUCGAUGAGCUGAUGCAGGCCCUGAAUUUCAACAGCGAGACCGUGCCCCAAAAAAGCAGCCUGGAAGAACCGGAUUUUUAUAAAACCAAAAUCAAGCUGUGCAUACUGCUGCAUGCCUUCAGAAUCAGAGCCGUGACCAUCGAUAGAGUGAUGAGCUAUCUGAAUGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 156 hIL12AB_016G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAGCAGCUGGUCAUCAGCUGGUUCAGCCUGGUCUUCCUGGCCAGCCCCCUGGUG T100 tail)GCCAUCUGGGAGCUGAAGAAGGACGUAUACGUAGUGGAGUUGGAUUGGUACCCAGACGCUCCUGGGGAGAUGGUGGUGCUGACCUGUGACACCCCAGAAGAGGACGGUAUCACCUGGACCCUGGACCAGAGCUCAGAAGUGCUGGGCAGUGGAAAAACCCUGACCAUCCAGGUGAAGGAGUUUGGAGAUGCUGGCCAGUACACCUGCCACAAGGGUGGUGAAGUGCUGAGCCACAGCCUGCUGCUGCUGCACAAGAAGGAGGAUGGCAUCUGGAGCACAGAUAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACCUUCCUUCGCUGUGAAGCCAAGAACUACAGUGGCCGCUUCACCUGCUGGUGGCUGACCACCAUCAGCACAGACCUCACCUUCUCGGUGAAGAGCAGCAGAGGCAGCUCAGACCCCCAGGGUGUCACCUGUGGGGCGGCCACGCUGUCGGCGGAGAGAGUUCGAGGUGACAACAAGGAGUAUGAAUACUCGGUGGAGUGCCAGGAAGAUUCGGCGUGCCCGGCGGCAGAAGAGAGCCUGCCCAUAGAAGUGAUGGUGGAUGCUGUGCACAAGCUGAAGUAUGAAAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAGCCAGACCCGCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCCGGCAGGUGGAGGUUUCCUGGGAGUACCCAGAUACGUGGAGCACCCCCCACAGCUACUUCAGCCUGACCUUCUGUGUCCAGGUGCAGGGCAAGAGCAAGAGAGAGAAGAAAGAUAGAGUCUUCACAGAUAAGACCUCGGCCACGGUCAUCUGCAGAAAGAAUGCCUCCAUCUCGGUUCGAGCCCAAGAUAGAUACUACAGCAGCAGCUGGUCAGAAUGGGCCUCGGUGCCCUGCAGUGGUGGCGGCGGCGGCGGCAGCAGAAACCUGCCUGUUGCCACCCCAGACCCUGGGAUGUUCCCCUGCCUGCACCACAGCCAGAACUUAUUACGAGCUGUUUCUAACAUGCUGCAGAAGGCCCGGCAGACCCUGGAGUUCUACCCCUGCACCUCAGAAGAGAUUGACCAUGAAGAUAUCACCAAAGAUAAGACCAGCACUGUAGAGGCCUGCCUGCCCCUGGAGCUGACCAAGAAUGAAAGCUGCCUGAACAGCAGAGAGACCAGCUUCAUCACCAAUGGAAGCUGCCUGGCCAGCAGAAAGACCAGCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUAUGAAGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAAUGCAAAGCUGCUGAUGGACCCCAAGCGGCAGAUAUUUUUGGACCAGAACAUGCUGGCUGUCAUUGAUGAGCUGAUGCAGGCCCUGAACUUCAACUCAGAAACUGUACCCCAGAAGAGCAGCCUGGAGGAGCCAGAUUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUUCAUGCUUUCAGAAUCAGAGCUGUCACCAUUGACCGCGUGAUGAGCUACUUAAAUGCCUCGUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 157 hIL12AB_017G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAGCAGCUGGUAAUCAGCUGGUUUUCCCUCGUCUUUCUGGCAUCACCCCUGGUG T100 tail)GCUAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGCUGGAUUGGUACCCUGACGCCCCGGGGGAAAUGGUGGUGUUAACCUGCGACACGCCUGAGGAGGACGGCAUCACCUGGACGCUGGACCAGAGCAGCGAGGUGCUUGGGUCUGGUAAAACUCUGACUAUUCAGGUGAAAGAGUUCGGGGAUGCCGGCCAAUAUACUUGCCACAAGGGUGGCGAGGUGCUUUCUCAUUCUCUGCUCCUGCUGCACAAGAAAGAAGAUGGCAUUUGGUCUACUGAUAUUCUGAAAGACCAGAAGGAGCCCAAGAACAAGACCUUUCUGAGAUGCGAGGCUAAAAACUACAGCGGAAGAUUUACCUGCUGGUGGCUGACCACAAUCUCAACCGACCUGACAUUUUCAGUGAAGUCCAGCAGAGGGAGCUCCGACCCUCAGGGCGUGACCUGCGGAGCCGCCACUCUGUCCGCAGAAAGAGUGAGAGGUGAUAAUAAGGAGUACGAGUAUUCAGUCGAGUGCCAAGAAGAUUCUGCCUGCCCAGCCGCCGAGGAGAGCCUGCCAAUCGAGGUGAUGGUAGAUGCGGUACACAAGCUGAAGUAUGAGAACUACACAUCCUCCUUCUUCAUAAGAGAUAUUAUCAAGCCUGACCCACCUAAAAAUCUGCAACUCAAGCCUUUGAAAAAUUCACGGCAGGUGGAGGUGAGCUGGGAGUACCCUGAUACUUGGAGCACCCCCCAUAGCUACUUUUCGCUGACAUUCUGCGUCCAGGUGCAGGGCAAGUCAAAGAGAGAGAAGAAGGAUCGCGUGUUCACUGAUAAAACAAGCGCCACAGUGAUCUGCAGAAAAAACGCUAGCAUUAGCGUCAGAGCACAGGACCGGUAUUACUCCAGCUCCUGGAGCGAAUGGGCAUCUGUGCCCUGCAGCGGUGGGGGCGGAGGCGGAUCCAGAAACCUCCCCGUUGCCACACCUGAUCCUGGAAUGUUCCCCUGUCUGCACCACAGCCAGAACCUGCUGAGAGCAGUGUCUAACAUGCUCCAGAAGGCCAGGCAGACCCUGGAGUUUUACCCCUGCACCAGCGAGGAAAUCGAUCACGAAGAUAUCACCAAAGAUAAAACCUCCACCGUGGAGGCCUGCCUGCCCCUGGAACUGACCAAAAACGAGAGCUGCCUGAAUAGCAGGGAGACCUCCUUCAUCACCAACGGCUCAUGCCUUGCCAGCCGGAAAACUAGCUUCAUGAUGGCCCUGUGCCUGUCUUCGAUCUAUGAGGACCUGAAAAUGUACCAGGUCGAAUUUAAGACGAUGAACGCAAAGCUGCUGAUGGACCCCAAGCGGCAGAUCUUUCUGGACCAGAACAUGCUGGCAGUCAUAGAUGAGUUGAUGCAGGCAUUAAACUUCAACAGCGAGACCGUGCCUCAGAAGUCCAGCCUCGAGGAGCCAGAUUUUUAUAAGACCAAGAUCAAACUAUGCAUCCUGCUGCAUGCUUUCAGGAUUAGAGCCGUCACCAUCGAUCGAGUCAUGUCUUACCUGAAUGCUAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 158 hIL12AB_018G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGUC (mRNA withACCAACAGUUAGUAAUCUCCUGGUUUUCUCUGGUGUUUCUGGCCAGCCCCCUCGUG T100 tail)GCCAUCUGGGAGCUUAAAAAGGACGUUUACGUGGUGGAGUUGGAUUGGUAUCCCGACGCUCCAGGCGAAAUGGUCGUGCUGACCUGCGAUACCCCUGAAGAAGACGGUAUCACCUGGACGCUGGACCAGUCUUCCGAGGUGCUUGGAUCUGGCAAAACACUGACAAUACAAGUUAAGGAGUUCGGGGACGCAGGGCAGUACACCUGCCACAAAGGCGGCGAGGUCCUGAGUCACUCCCUGUUACUGCUCCACAAGAAAGAGGACGGCAUUUGGUCCACCGACAUUCUGAAGGACCAGAAGGAGCCUAAGAAUAAAACUUUCCUGAGAUGCGAGGCAAAAAACUAUAGCGGCCGCUUUACUUGCUGGUGGCUUACAACAAUCUCUACCGAUUUAACUUUCUCCGUGAAGUCUAGCAGAGGAUCCUCUGACCCGCAAGGAGUGACUUGCGGAGCCGCCACCUUGAGCGCCGAAAGAGUCCGUGGCGAUAACAAAGAAUACGAGUACUCCGUGGAGUGCCAGGAAGAUUCCGCCUGCCCAGCUGCCGAGGAGUCCCUGCCCAUUGAAGUGAUGGUGGAUGCCGUCCACAAGCUGAAGUACGAAAACUAUACCAGCAGCUUCUUCAUCCGGGAUAUCAUUAAGCCCGACCCUCCUAAAAACCUGCAACUUAAGCCCCUAAAGAAUAGUCGGCAGGUUGAGGUCAGCUGGGAAUAUCCUGACACAUGGAGCACCCCCCACUCUUAUUUCUCCCUGACCUUCUGCGUGCAGGUGCAGGGCAAGAGUAAACGGGAGAAAAAAGAUAGGGUCUUUACCGAUAAAACCAGCGCUACGGUUAUCUGUCGGAAGAACGCUUCCAUCUCCGUCCGCGCUCAGGAUCGUUACUACUCGUCCUCAUGGAGCGAGUGGGCCAGCGUGCCCUGCAGCGGCGGCGGUGGAGGCGGAUCCAGAAAUCUGCCUGUUGCCACACCAGACCCUGGCAUGUUCCCCUGUCUGCAUCAUAGCCAGAACCUGCUCAGAGCCGUGAGCAACAUGCUCCAGAAGGCCAGGCAAACUUUGGAGUUCUACCCGUGUACAUCUGAGGAAAUCGAUCACGAAGAUAUAACCAAAGAUAAAACCUCUACAGUAGAGGCUUGUUUGCCCCUGGAGUUGACCAAAAACGAGAGUUGCCUGAACAGUCGCGAGACGAGCUUCAUUACUAACGGCAGCUGUCUCGCCUCCAGAAAAACAUCCUUCAUGAUGGCCCUGUGUCUUUCCAGCAUAUACGAAGACCUGAAAAUGUACCAGGUCGAGUUCAAAACAAUGAACGCCAAGCUGCUUAUGGACCCCAAGCGGCAGAUCUUCCUCGACCAAAACAUGCUCGCUGUGAUCGAUGAGCUGAUGCAGGCUCUCAACUUCAAUUCCGAAACAGUGCCACAGAAGUCCAGUCUGGAAGAACCCGACUUCUACAAGACCAAGAUUAAGCUGUGUAUUUUGCUGCAUGCGUUUAGAAUCAGAGCCGUGACCAUUGAUCGGGUGAUGAGCUACCUGAACGCCUCGUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 159 hIL12AB_019G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAGCAGCUUGUCAUCUCCUGGUUUUCUCUUGUCUUCCUGGCCUCGCCGCUGGUG T100 tail)GCCAUCUGGGAGCUGAAGAAAGACGUUUACGUAGUAGAGUUGGAUUGGUACCCAGACGCACCUGGAGAAAUGGUGGUUCUCACCUGUGACACUCCUGAAGAAGACGGUAUCACCUGGACGCUGGACCAAAGCUCAGAAGUUCUUGGCAGUGGAAAAACGCUGACCAUACAAGUAAAAGAAUUUGGGGAUGCUGGCCAGUACACGUGCCACAAAGGAGGAGAAGUUCUCAGCCACAGUUUACUUCUUCUUCACAAGAAAGAAGAUGGCAUCUGGUCCACAGAUAUUUUAAAAGACCAGAAGGAGCCCAAGAACAAAACCUUCCUCCGCUGUGAGGCCAAGAACUACAGUGGUCGUUUCACCUGCUGGUGGCUCACCACCAUCUCCACUGACCUCACCUUCUCUGUAAAAAGCAGCCGUGGUUCUUCUGACCCCCAAGGAGUCACCUGUGGGGCUGCCACGCUCUCGGCAGAAAGAGUUCGAGGUGACAACAAGGAAUAUGAAUAUUCUGUGGAAUGUCAAGAAGAUUCUGCCUGCCCGGCGGCAGAAGAAAGUCUUCCCAUAGAAGUCAUGGUGGAUGCUGUUCACAAAUUAAAAUAUGAAAACUACACCAGCAGCUUCUUCAUUCGUGACAUCAUCAAACCAGACCCGCCCAAGAACCUUCAGUUAAAACCUUUAAAAAACAGCCGGCAGGUAGAAGUUUCCUGGGAGUACCCAGAUACGUGGUCCACGCCGCACUCCUACUUCAGUUUAACCUUCUGUGUACAAGUACAAGGAAAAUCAAAAAGAGAGAAGAAAGAUCGUGUCUUCACUGACAAAACAUCUGCCACGGUCAUCUGCAGGAAGAAUGCCUCCAUCUCGGUUCGAGCCCAGGACCGCUACUACAGCAGCAGCUGGAGUGAGUGGGCAUCUGUUCCCUGCAGUGGUGGCGGCGGCGGCGGCAGCCGCAACCUUCCUGUGGCCACGCCGGACCCUGGCAUGUUCCCGUGCCUUCACCACUCCCAAAAUCUUCUUCGUGCUGUUUCUAACAUGCUGCAGAAGGCGCGCCAAACUUUAGAAUUCUACCCGUGCACUUCUGAAGAAAUAGACCAUGAAGAUAUCACCAAAGAUAAAACCAGCACGGUGGAGGCCUGCCUUCCUUUAGAGCUGACCAAGAAUGAAUCCUGCCUCAACAGCAGAGAGACCAGCUUCAUCACCAAUGGCAGCUGCCUGGCCUCGCGCAAGACCAGCUUCAUGAUGGCGCUGUGCCUUUCUUCCAUCUAUGAAGAUUUAAAGAUGUACCAAGUAGAAUUUAAAACCAUGAAUGCCAAAUUAUUAAUGGACCCCAAACGGCAGAUAUUUUUGGAUCAAAACAUGCUGGCUGUCAUUGAUGAGCUCAUGCAAGCAUUAAACUUCAACUCAGAAACUGUUCCCCAGAAGUCAUCUUUAGAAGAGCCAGAUUUCUACAAAACAAAAAUAAAACUCUGCAUUCUUCUUCAUGCCUUCCGCAUCCGUGCUGUCACCAUUGACCGUGUCAUGUCCUACUUAAAUGCUUCUUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 160 hIL12AB_020G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAGCAGCUGGUGAUCAGCUGGUUCAGCCUGGUGUUCCUGGCUAGCCCUCUGGUG T100 tail)GCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGUUGGAUUGGUACCCCGACGCUCCCGGCGAGAUGGUGGUGCUGACCUGCGACACCCCCGAGGAGGACGGGAUCACCUGGACCCUGGAUCAGUCAAGCGAGGUGCUGGGAAGCGGCAAGACCCUGACCAUCCAGGUGAAGGAGUUCGGCGACGCCGGCCAAUACACUUGCCACAAGGGAGGCGAGGUGCUGUCCCACUCCCUCCUGCUGCUGCACAAAAAGGAAGACGGCAUCUGGAGCACCGACAUCCUGAAAGACCAGAAGGAGCCUAAGAACAAAACAUUCCUCAGAUGCGAGGCCAAGAAUUACUCCGGGAGAUUCACCUGUUGGUGGCUGACCACCAUCAGCACAGACCUGACCUUCAGCGUGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGUGACCUGUGGCGCCGCCACCCUGAGCGCCGAAAGAGUGCGCGGCGACAACAAGGAGUACGAGUACUCCGUGGAAUGCCAGGAAGAUAGCGCCUGCCCCGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUCCACAAGCUGAAGUACGAGAACUACACCUCUAGCUUCUUCAUCAGAGAUAUCAUCAAGCCCGAUCCCCCCAAGAACCUGCAGCUGAAACCCCUGAAGAACAGCCGGCAGGUGGAGGUGAGCUGGGAGUAUCCCGACACCUGGUCCACCCCCCACAGCUAUUUUAGCCUGACCUUCUGCGUGCAAGUGCAGGGCAAGAGCAAGAGAGAGAAGAAGGACCGCGUGUUCACCGACAAAACCAGCGCCACCGUGAUCUGCAGAAAGAACGCCAGCAUCAGCGUGAGGGCCCAGGAUAGAUACUACAGUUCCAGCUGGAGCGAGUGGGCCAGCGUGCCCUGCAGCGGCGGCGGCGGGGGAGGCUCGAGAAACCUGCCCGUGGCUACCCCCGAUCCCGGAAUGUUCCCCUGCCUGCACCACAGCCAGAACCUGCUGAGGGCGGUGUCCAACAUGCUUCAGAAGGCCCGGCAGACCCUGGAGUUCUACCCCUGUACCUCUGAGGAGAUCGAUCAUGAAGAUAUCACAAAAGAUAAAACCAGCACCGUGGAGGCCUGCCUGCCCCUGGAGCUGACCAAGAACGAGAGCUGCCUGAACUCCCGCGAGACCAGCUUCAUCACGAACGGCAGCUGCCUGGCCAGCAGGAAGACCUCCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAAAUGUACCAGGUGGAGUUUAAGACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGGCAAAUCUUCCUGGACCAGAACAUGCUGGCAGUGAUCGACGAGCUCAUGCAGGCCCUGAACUUCAAUAGCGAGACGGUCCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUUUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUUAGAAUCCGUGCCGUGACCAUUGACAGAGUGAUGAGCUACCUGAAUGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 161 hIL12AB_021G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAGCAGCUGGUGAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCAGCCCUCUGGUU T100 tail)GCCAUCUGGGAGCUGAAGAAAGACGUGUACGUCGUGGAACUGGACUGGUAUCCGGACGCCCCGGGCGAGAUGGUGGUGCUGACCUGUGACACCCCCGAGGAGGACGGCAUCACCUGGACGCUGGACCAAUCCUCCGAGGUGCUGGGAAGCGGCAAGACCCUGACCAUCCAGGUGAAGGAAUUCGGGGACGCCGGGCAGUACACCUGCCACAAGGGGGGCGAAGUGCUGUCCCACUCGCUGCUGCUCCUGCAUAAGAAGGAGGAUGGAAUCUGGUCCACCGACAUCCUCAAAGAUCAGAAGGAGCCCAAGAACAAGACGUUCCUGCGCUGUGAAGCCAAGAAUUAUUCGGGGCGAUUCACGUGCUGGUGGCUGACAACCAUCAGCACCGACCUGACGUUUAGCGUGAAGAGCAGCAGGGGGUCCAGCGACCCCCAGGGCGUGACGUGCGGCGCCGCCACCCUCUCCGCCGAGAGGGUGCGGGGGGACAAUAAGGAGUACGAGUACAGCGUGGAAUGCCAGGAGGACAGCGCCUGCCCCGCCGCGGAGGAAAGCCUCCCGAUAGAGGUGAUGGUGGACGCCGUGCACAAGCUCAAGUAUGAGAAUUACACCAGCAGCUUUUUCAUCCGGGACAUUAUCAAGCCCGACCCCCCGAAGAACCUCCAGCUGAAGCCCCUGAAGAACAGCCGGCAGGUGGAAGUCUCCUGGGAGUAUCCCGACACCUGGAGCACCCCGCACAGCUACUUCUCCCUGACCUUCUGUGUGCAGGUGCAGGGCAAGUCCAAGAGGGAAAAGAAGGACAGGGUUUUCACCGACAAGACCAGCGCGACCGUGAUCUGCCGGAAGAACGCCAGCAUAAGCGUCCGCGCCCAAGAUAGGUACUACAGCAGCUCCUGGAGCGAGUGGGCUAGCGUGCCCUGCAGCGGGGGCGGGGGUGGGGGCUCCAGGAACCUGCCAGUGGCGACCCCCGACCCCGGCAUGUUCCCCUGCCUCCAUCACAGCCAGAACCUGCUGAGGGCCGUCAGCAAUAUGCUGCAGAAGGCCAGGCAGACCCUGGAAUUCUACCCCUGCACGUCGGAGGAGAUCGAUCACGAGGAUAUCACAAAAGACAAGACUUCCACCGUGGAGGCCUGCCUGCCCCUGGAGCUCACCAAGAAUGAGUCCUGUCUGAACUCCCGGGAAACCAGCUUCAUCACCAACGGGUCCUGCCUGGCCAGCAGGAAGACCAGCUUUAUGAUGGCCCUGUGCCUGUCGAGCAUCUACGAGGACCUGAAGAUGUACCAGGUCGAGUUCAAGACAAUGAACGCCAAGCUGCUGAUGGACCCCAAGAGGCAAAUCUUCCUGGACCAGAAUAUGCUUGCCGUCAUCGACGAGCUCAUGCAGGCCCUGAACUUCAACUCCGAGACCGUGCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCGUUCAGGAUCCGGGCAGUCACCAUCGACCGUGUGAUGUCCUACCUGAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 162 hIL12AB_022G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withAUCAGCAGCUGGUGAUCAGCUGGUUCAGCCUGGUGUUCCUCGCCUCUCCCCUGGUG T100 tail)GCCAUCUGGGAGCUCAAAAAGGACGUGUACGUGGUGGAGCUCGACUGGUACCCAGACGCCCCCGGGGAGAUGGUGGUGCUGACCUGCGACACCCCCGAAGAAGACGGCAUCACGUGGACCCUCGACCAGUCCAGCGAGGUGCUGGGGAGCGGGAAGACUCUGACCAUCCAGGUCAAGGAGUUCGGGGACGCCGGGCAGUACACGUGCCACAAGGGCGGCGAAGUCUUAAGCCACAGCCUGCUCCUGCUGCACAAGAAGGAGGACGGGAUCUGGUCCACAGACAUACUGAAGGACCAGAAGGAGCCGAAGAAUAAAACCUUUCUGAGGUGCGAGGCCAAGAACUAUUCCGGCAGGUUCACGUGCUGGUGGCUUACAACAAUCAGCACAGACCUGACGUUCAGCGUGAAGUCCAGCCGCGGCAGCAGCGACCCCCAGGGGGUGACCUGCGGCGCCGCCACCCUGAGCGCCGAGCGGGUGCGCGGGGACAACAAGGAGUACGAGUACUCCGUGGAGUGCCAGGAAGACAGCGCCUGUCCCGCCGCCGAAGAGAGCCUGCCUAUCGAGGUCAUGGUAGAUGCAGUGCAUAAGCUGAAGUACGAGAACUAUACGAGCAGCUUUUUCAUACGCGACAUCAUCAAGCCCGACCCCCCCAAGAACCUGCAGCUUAAGCCCCUGAAGAAUAGCCGGCAGGUGGAGGUCUCCUGGGAGUACCCCGACACCUGGUCAACGCCCCACAGCUACUUCUCCCUGACCUUUUGUGUCCAAGUCCAGGGAAAGAGCAAGAGGGAGAAGAAAGAUCGGGUGUUCACCGACAAGACCUCCGCCACGGUGAUCUGCAGGAAGAACGCCAGCAUCUCCGUGAGGGCGCAAGACAGGUACUACUCCAGCAGCUGGUCCGAAUGGGCCAGCGUGCCCUGCUCCGGCGGCGGGGGCGGCGGCAGCCGAAACCUACCCGUGGCCACGCCGGAUCCCGGCAUGUUUCCCUGCCUGCACCACAGCCAGAACCUCCUGAGGGCCGUGUCCAACAUGCUGCAGAAGGCCAGGCAGACUCUGGAGUUCUACCCCUGCACGAGCGAGGAGAUCGAUCACGAGGACAUCACCAAGGAUAAGACCAGCACUGUGGAGGCCUGCCUUCCCCUGGAGCUGACCAAGAACGAGAGCUGUCUGAACUCCAGGGAGACCUCAUUCAUCACCAACGGCUCCUGCCUGGCCAGCAGGAAAACCAGCUUCAUGAUGGCCUUGUGUCUCAGCUCCAUCUACGAGGACCUGAAGAUGUAUCAGGUCGAGUUCAAGACAAUGAACGCCAAGCUGCUGAUGGACCCCAAAAGGCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUCAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAGACGGUGCCCCAGAAAAGCUCCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCAGGAUCAGGGCAGUGACCAUCGACCGGGUGAUGUCAUACCUUAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 163 hIL12AB_023G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withAUCAGCAGCUGGUGAUCUCCUGGUUCAGCCUGGUGUUUCUGGCCUCGCCCCUGGUC T100 tail)GCCAUCUGGGAGCUGAAGAAAGACGUGUACGUCGUCGAACUGGACUGGUACCCCGACGCCCCCGGGGAGAUGGUGGUGCUGACCUGCGACACGCCGGAGGAGGACGGCAUCACCUGGACCCUGGAUCAAAGCAGCGAGGUGCUGGGCAGCGGCAAGACCCUGACCAUCCAAGUGAAGGAAUUCGGCGAUGCCGGCCAGUACACCUGUCACAAAGGGGGCGAGGUGCUCAGCCACAGCCUGCUGCUGCUGCACAAGAAGGAGGAUGGCAUCUGGAGCACCGAUAUCCUGAAGGACCAGAAAGAGCCCAAGAACAAGACGUUCCUGAGGUGCGAGGCCAAGAACUACAGCGGUAGGUUCACGUGUUGGUGGCUGACCACCAUCAGCACCGACCUGACGUUCAGCGUGAAGAGCUCCAGGGGCAGCUCCGACCCACAGGGGGUGACGUGCGGGGCCGCAACCCUCAGCGCCGAAAGGGUGCGGGGGGACAACAAGGAGUACGAAUACUCCGUGGAGUGCCAGGAAGAUUCGGCCUGCCCCGCCGCGGAGGAGAGCCUCCCCAUCGAGGUAAUGGUGGACGCCGUGCAUAAGCUGAAGUACGAGAACUACACCAGCUCGUUCUUCAUCCGAGACAUCAUCAAACCCGACCCGCCCAAAAAUCUGCAGCUCAAGCCCCUGAAGAACUCCAGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGUCCACCCCGCACAGCUACUUCUCCCUGACAUUCUGCGUGCAGGUGCAGGGCAAGAGCAAGCGGGAGAAGAAGGACAGGGUGUUCACCGACAAGACGAGCGCCACCGUGAUCUGCCGAAAGAACGCCAGCAUCUCGGUGCGCGCCCAGGAUAGGUACUAUUCCAGCUCCUGGAGCGAGUGGGCCUCGGUACCCUGCAGCGGCGGCGGGGGCGGCGGCAGUAGGAAUCUGCCCGUGGCUACCCCGGACCCGGGCAUGUUCCCCUGCCUCCACCACAGCCAGAACCUGCUGAGGGCCGUGAGCAACAUGCUGCAGAAGGCCAGACAGACGCUGGAGUUCUACCCCUGCACGAGCGAGGAGAUCGACCACGAGGACAUCACCAAGGAUAAAACUUCCACCGUCGAGGCCUGCCUGCCCUUGGAGCUGACCAAGAAUGAAUCCUGUCUGAACAGCAGGGAGACCUCGUUUAUCACCAAUGGCAGCUGCCUCGCCUCCAGGAAGACCAGCUUCAUGAUGGCCCUCUGUCUGAGCUCCAUCUAUGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCGAAGCUGCUGAUGGACCCCAAGAGGCAGAUCUUCCUGGAUCAGAAUAUGCUGGCGGUGAUCGACGAGCUCAUGCAGGCCCUCAAUUUCAAUAGCGAGACAGUGCCCCAGAAGUCCUCCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGUAUCCUGCUGCACGCCUUCCGGAUCCGGGCCGUCACCAUCGACCGGGUCAUGAGCUACCUCAAUGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 164 hIL12AB_024G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAGCAGCUGGUGAUCUCCUGGUUCUCCCUGGUGUUCCUGGCCUCGCCCCUGGUG T100 tail)GCCAUCUGGGAGCUGAAGAAGGACGUGUACGUCGUGGAGCUCGACUGGUACCCCGACGCCCCUGGCGAGAUGGUGGUGCUGACCUGCGACACCCCAGAGGAGGAUGGCAUCACCUGGACCCUGGAUCAGUCCUCCGAGGUGCUGGGCUCCGGCAAGACGCUGACCAUCCAAGUGAAGGAGUUCGGUGACGCCGGACAGUAUACCUGCCAUAAGGGCGGCGAGGUCCUGUCCCACAGCCUCCUCCUCCUGCAUAAGAAGGAGGACGGCAUCUGGAGCACCGACAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACCUUUCUGAGGUGCGAGGCCAAGAACUACAGCGGCCGAUUCACCUGCUGGUGGCUCACCACCAUAUCCACCGACCUGACUUUCUCCGUCAAGUCCUCCCGGGGGUCCAGCGACCCCCAGGGAGUGACCUGCGGCGCCGCCACCCUCAGCGCCGAGCGGGUGCGGGGGGACAACAAGGAGUACGAAUACUCCGUCGAGUGCCAGGAGGACUCCGCCUGCCCGGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUCGACGCGGUGCACAAGCUGAAGUACGAGAACUACACCAGCAGUUUCUUCAUCAGGGAUAUCAUCAAGCCAGAUCCCCCGAAGAAUCUGCAACUGAAGCCGCUGAAAAACUCACGACAGGUGGAGGUGAGCUGGGAGUACCCCGACACGUGGAGCACCCCACAUUCCUACUUCAGCCUGACCUUCUGCGUGCAGGUCCAGGGCAAGAGCAAGCGGGAGAAGAAGGACAGGGUGUUCACGGAUAAGACCAGUGCCACCGUGAUCUGCAGGAAGAACGCCUCUAUUAGCGUGAGGGCCCAGGAUCGGUAUUACUCCUCGAGCUGGAGCGAAUGGGCCUCCGUGCCCUGCAGUGGGGGGGGUGGAGGCGGGAGCAGGAACCUGCCCGUAGCAACCCCCGACCCCGGGAUGUUCCCCUGUCUGCACCACUCGCAGAACCUGCUGCGCGCGGUGAGCAACAUGCUCCAAAAAGCCCGUCAGACCUUAGAGUUCUACCCCUGCACCAGCGAAGAAAUCGACCACGAAGACAUCACCAAGGACAAAACCAGCACCGUGGAGGCGUGCCUGCCGCUGGAGCUGACCAAGAACGAGAGCUGCCUCAACUCCAGGGAGACCAGCUUUAUCACCAACGGCUCGUGCCUAGCCAGCCGGAAAACCAGCUUCAUGAUGGCCCUGUGCCUGAGCUCCAUUUACGAGGACCUGAAGAUGUAUCAGGUGGAGUUCAAGACCAUGAAUGCCAAACUCCUGAUGGACCCCAAGAGGCAGAUCUUCCUGGACCAGAACAUGCUCGCGGUGAUCGAUGAGCUGAUGCAGGCCCUGAACUUUAAUAGCGAGACCGUGCCCCAGAAAAGCAGCCUGGAGGAGCCGGACUUCUACAAGACCAAAAUCAAGCUGUGCAUCCUGCUCCACGCCUUCCGCAUCCGGGCCGUGACCAUCGACAGGGUGAUGAGCUACCUGAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 165 hIL12AB_025G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withAUCAGCAGCUGGUGAUUUCCUGGUUCUCCCUGGUGUUCCUGGCCAGCCCCCUCGUG T100 tail)GCGAUCUGGGAGCUAAAGAAGGACGUGUACGUGGUGGAGCUGGACUGGUACCCGGACGCACCCGGCGAGAUGGUCGUUCUGACCUGCGAUACGCCAGAGGAGGACGGCAUCACCUGGACCCUCGAUCAGAGCAGCGAGGUCCUGGGGAGCGGAAAGACCCUGACCAUCCAGGUCAAGGAGUUCGGCGACGCCGGCCAGUACACCUGCCACAAAGGUGGCGAGGUCCUGAGCCACUCGCUGCUGCUCCUGCAUAAGAAGGAGGACGGAAUCUGGAGCACAGACAUCCUGAAAGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGAGGUGCGAGGCCAAGAACUACAGCGGGCGCUUCACGUGCUGGUGGCUGACCACCAUCAGCACGGACCUCACCUUCUCCGUGAAGAGCAGCCGGGGAUCCAGCGAUCCCCAAGGCGUCACCUGCGGCGCGGCCACCCUGAGCGCGGAGAGGGUCAGGGGCGAUAAUAAGGAGUAUGAGUACAGCGUGGAGUGCCAGGAGGACAGCGCCUGCCCGGCCGCCGAGGAGUCCCUGCCAAUCGAAGUGAUGGUCGACGCCGUGCACAAGCUGAAGUACGAGAACUACACCAGCAGCUUCUUCAUCCGGGAUAUCAUCAAGCCCGAUCCCCCGAAGAACCUGCAGCUGAAGCCCCUCAAGAACAGCCGGCAGGUGGAGGUGAGUUGGGAGUACCCCGACACCUGGUCAACGCCCCACAGCUACUUCUCCCUGACCUUCUGUGUGCAGGUGCAGGGAAAGAGCAAGAGGGAGAAGAAAGACCGGGUCUUCACCGACAAGACCAGCGCCACGGUGAUCUGCAGGAAGAACGCAAGCAUCUCCGUGAGGGCCCAGGACAGGUACUACAGCUCCAGCUGGUCCGAAUGGGCCAGCGUGCCCUGUAGCGGCGGCGGGGGCGGUGGCAGCCGCAACCUCCCAGUGGCCACCCCCGACCCCGGCAUGUUCCCCUGCCUGCACCACAGCCAGAAUCUGCUGAGGGCCGUGAGUAACAUGCUGCAGAAGGCAAGGCAAACCCUCGAAUUCUAUCCCUGCACCUCCGAGGAGAUCGACCACGAGGAUAUCACCAAGGACAAGACCAGCACCGUCGAGGCCUGUCUCCCCCUGGAGCUGACCAAGAAUGAGAGCUGCCUGAACAGCCGGGAGACCAGCUUCAUCACCAACGGGAGCUGCCUGGCCUCCAGGAAGACCUCGUUCAUGAUGGCGCUGUGCCUCUCAAGCAUAUACGAGGAUCUGAAGAUGUACCAGGUGGAGUUUAAGACGAUGAACGCCAAGCUGCUGAUGGACCCGAAGAGGCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUGAUAGACGAGCUCAUGCAGGCCCUGAACUUCAACUCCGAGACCGUGCCGCAGAAGUCAUCCCUCGAGGAGCCCGACUUCUAUAAGACCAAGAUCAAGCUGUGCAUCCUGCUCCACGCCUUCCGGAUAAGGGCCGUGACGAUCGACAGGGUGAUGAGCUACCUUAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 166 hIL12AB_026G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAGCAGCUCGUGAUCAGCUGGUUCUCCCUGGUGUUUCUCGCCAGCCCCCUGGUG T100 tail)GCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGCUGGACUGGUACCCUGACGCCCCGGGGGAGAUGGUCGUGCUGACCUGCGACACCCCCGAAGAGGACGGUAUCACCUGGACCCUGGACCAGUCCAGCGAGGUGCUGGGCAGCGGCAAGACCCUGACUAUUCAAGUCAAGGAGUUCGGAGACGCCGGCCAGUACACCUGCCACAAGGGUGGAGAGGUGUUAUCACACAGCCUGCUGCUGCUGCACAAGAAGGAAGACGGGAUCUGGAGCACCGACAUCCUGAAGGACCAGAAGGAGCCCAAAAACAAGACCUUCCUGCGGUGCGAGGCCAAGAACUAUUCGGGCCGCUUUACGUGCUGGUGGCUGACCACCAUCAGCACUGAUCUCACCUUCAGCGUGAAGUCCUCCCGGGGGUCGUCCGACCCCCAGGGGGUGACCUGCGGGGCCGCCACCCUGUCCGCCGAGAGAGUGAGGGGCGAUAAUAAGGAGUACGAGUACAGCGUUGAGUGCCAGGAAGAUAGCGCCUGUCCCGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUCCACAAGCUGAAGUAUGAGAACUACACCUCAAGCUUCUUCAUCAGGGACAUCAUCAAACCCGAUCCGCCCAAGAAUCUGCAGCUGAAGCCCCUGAAAAAUAGCAGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGUCCACCCCCCAUAGCUAUUUCUCCCUGACGUUCUGCGUGCAGGUGCAAGGGAAGAGCAAGCGGGAGAAGAAGGACCGGGUGUUCACCGACAAGACCUCCGCCACCGUGAUCUGUAGGAAGAACGCGUCGAUCUCGGUCAGGGCCCAGGACAGGUAUUACAGCAGCAGCUGGAGCGAGUGGGCGAGCGUGCCCUGCUCGGGCGGCGGCGGCGGCGGGAGCAGAAAUCUGCCCGUGGCCACCCCAGACCCCGGAAUGUUCCCCUGCCUGCACCAUUCGCAGAACCUCCUGAGGGCCGUGAGCAACAUGCUGCAGAAGGCCCGCCAGACGCUGGAGUUCUACCCCUGCACGAGCGAGGAGAUCGACCACGAAGACAUCACCAAGGACAAAACCAGCACCGUGGAGGCCUGCCUGCCCCUGGAGCUGACCAAAAACGAAUCCUGCCUCAACAGCCGGGAGACCAGCUUCAUCACCAACGGCAGCUGCCUGGCCAGCCGAAAGACCUCCUUCAUGAUGGCCCUCUGCCUGAGCAGCAUCUAUGAGGAUCUGAAGAUGUAUCAGGUGGAGUUCAAGACCAUGAAUGCCAAGCUGCUGAUGGACCCCAAGAGGCAGAUAUUCCUGGACCAGAAUAUGCUGGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAGACCGUCCCCCAGAAGUCCAGCCUGGAGGAGCCGGACUUUUACAAAACGAAGAUCAAGCUGUGCAUACUGCUGCACGCCUUCAGGAUCCGGGCCGUGACAAUCGACAGGGUGAUGUCCUACCUGAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 167 hIL12AB_027G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGUC (mRNA withACCAGCAGCUGGUGAUCAGCUGGUUCUCCCUGGUGUUCCUGGCCAGCCCCCUGGUG T100 tail)GCCAUCUGGGAGCUCAAGAAGGACGUCUACGUCGUGGAGCUGGAUUGGUACCCCGACGCUCCCGGGGAGAUGGUGGUGCUGACCUGCGACACCCCCGAGGAGGACGGCAUCACCUGGACGCUGGACCAGAGCUCAGAGGUGCUGGGAAGCGGAAAGACACUGACCAUCCAGGUGAAGGAGUUCGGGGAUGCCGGGCAGUAUACCUGCCACAAGGGCGGCGAAGUGCUGAGCCAUUCCCUGCUGCUGCUGCACAAGAAGGAGGACGGCAUAUGGUCCACCGACAUCCUGAAGGAUCAGAAGGAGCCGAAGAAUAAAACCUUCCUGAGGUGCGAGGCCAAGAAUUACAGCGGCCGAUUCACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUGACCUUCAGUGUGAAGUCCUCACGGGGCAGCUCAGAUCCCCAGGGCGUGACCUGCGGGGCCGCGACACUCAGCGCCGAGCGGGUGAGGGGUGAUAACAAGGAGUACGAGUAUUCUGUGGAGUGCCAGGAAGACUCCGCCUGUCCCGCCGCCGAGGAGUCCCUGCCCAUCGAGGUGAUGGUGGACGCCGUGCAUAAACUGAAGUACGAGAACUACACCUCCAGCUUCUUCAUCCGGGAUAUAAUCAAGCCCGACCCUCCGAAAAACCUGCAGCUGAAGCCCCUUAAAAACAGCCGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGAGCACCCCCCAUAGCUAUUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGGAAGUCCAAGCGCGAGAAAAAGGACCGGGUGUUCACCGACAAGACGAGCGCCACCGUGAUCUGCCGGAAGAACGCCAGUAUAAGCGUAAGGGCCCAGGAUAGGUACUACAGCUCCAGCUGGUCGGAGUGGGCCUCCGUGCCCUGUUCCGGCGGCGGGGGGGGUGGCAGCAGGAACCUCCCCGUGGCCACGCCGGACCCCGGCAUGUUCCCGUGCCUGCACCACUCCCAAAACCUCCUGCGGGCCGUCAGCAACAUGCUGCAAAAGGCGCGGCAGACCCUGGAGUUUUACCCCUGUACCUCCGAAGAGAUCGACCACGAGGAUAUCACCAAGGAUAAGACCUCCACCGUGGAGGCCUGUCUCCCCCUGGAGCUGACCAAGAACGAGAGCUGUCUUAACAGCAGAGAGACCUCGUUCAUAACGAACGGCUCCUGCCUCGCUUCCAGGAAGACGUCGUUCAUGAUGGCGCUGUGCCUGUCCAGCAUCUACGAGGACCUGAAGAUGUAUCAGGUCGAGUUCAAAACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGAGGCAGAUCUUCCUGGACCAGAACAUGCUCGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAAACCGUGCCCCAGAAGUCAAGCCUGGAGGAGCCGGACUUCUAUAAGACCAAGAUCAAGCUGUGUAUCCUGCUACACGCUUUUCGUAUCCGGGCCGUGACCAUCGACAGGGUUAUGUCGUACUUGAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 168 hIL12AB_028G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAACAGCUCGUGAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCAGCCCGCUGGUG T100 tail)GCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGCUGGACUGGUACCCCGACGCCCCCGGCGAGAUGGUGGUCCUGACCUGCGACACGCCGGAAGAGGACGGCAUCACCUGGACCCUGGAUCAGUCCAGCGAGGUGCUGGGCUCCGGCAAGACCCUGACCAUUCAGGUGAAGGAGUUCGGCGACGCCGGUCAGUACACCUGCCACAAGGGCGGCGAGGUGCUGAGCCACAGCCUACUGCUCCUGCACAAAAAGGAGGAUGGAAUCUGGUCCACCGACAUCCUCAAGGACCAGAAGGAGCCGAAGAACAAGACGUUCCUCCGGUGCGAGGCCAAGAACUACAGCGGCAGGUUUACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUGACAUUUUCCGUGAAGAGCAGCCGCGGCAGCAGCGAUCCCCAGGGCGUGACCUGCGGGGCGGCCACCCUGUCCGCCGAGCGUGUGAGGGGCGACAACAAGGAGUACGAGUACAGCGUGGAAUGCCAGGAGGACAGCGCCUGUCCCGCCGCCGAGGAGAGCCUGCCAAUCGAGGUCAUGGUGGACGCCGUGCACAAGCUGAAGUACGAGAACUACACGAGCAGCUUCUUCAUCAGGGACAUCAUCAAACCGGACCCGCCCAAGAACCUGCAGCUGAAACCCUUGAAAAACAGCAGGCAGGUGGAAGUGUCUUGGGAGUACCCCGACACCUGGUCCACCCCCCACAGCUACUUUAGCCUGACCUUCUGUGUGCAGGUCCAGGGCAAGUCCAAGAGGGAGAAGAAGGACAGGGUGUUCACCGACAAAACCAGCGCCACCGUGAUCUGCAGGAAGAACGCCUCCAUCAGCGUGCGGGCCCAGGACAGGUAUUACAGCUCGUCGUGGAGCGAGUGGGCCAGCGUGCCCUGCUCCGGGGGAGGCGGCGGCGGAAGCCGGAAUCUGCCCGUGGCCACCCCCGAUCCCGGCAUGUUCCCGUGUCUGCACCACAGCCAGAACCUGCUGCGGGCCGUGAGCAACAUGCUGCAGAAGGCCCGCCAAACCCUGGAGUUCUACCCCUGUACAAGCGAGGAGAUCGACCAUGAGGACAUUACCAAGGACAAGACCAGCACCGUGGAGGCCUGCCUGCCCCUCGAGCUCACAAAGAACGAAUCCUGCCUGAAUAGCCGCGAGACCAGCUUUAUCACGAACGGGUCCUGCCUCGCCAGCCGGAAGACAAGCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAAAUGUACCAAGUGGAGUUCAAAACGAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGCCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUCAUCGACGAGCUCAUGCAGGCCCUGAACUUCAACAGCGAGACCGUGCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUCUACAAGACGAAGAUCAAGCUCUGCAUCCUGCUGCACGCUUUCCGCAUCCGCGCGGUGACCAUCGACCGGGUGAUGAGCUACCUCAACGCCAGUUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 169 hIL12AB_029G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAACAGCUGGUGAUCAGCUGGUUCAGCCUGGUGUUUCUGGCCUCCCCUCUGGUG T100 tail)GCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGCUGGACUGGUACCCUGACGCCCCCGGCGAAAUGGUGGUGCUGACGUGCGACACCCCCGAGGAGGAUGGCAUCACCUGGACCCUGGACCAAAGCAGCGAGGUCCUCGGAAGCGGCAAGACCCUCACUAUCCAAGUGAAGGAGUUCGGGGAUGCGGGCCAGUACACCUGCCACAAGGGCGGCGAGGUGCUGUCUCAUAGCCUGCUGCUCCUGCAUAAGAAGGAAGACGGCAUCUGGAGCACCGACAUACUGAAGGAUCAGAAGGAGCCCAAGAACAAGACCUUCCUGAGGUGCGAGGCCAAGAACUACUCCGGGCGCUUCACCUGUUGGUGGCUGACCACCAUCUCCACCGACCUGACCUUCAGCGUGAAGAGCAGCAGGGGGAGCAGCGACCCCCAGGGGGUGACCUGCGGAGCCGCGACCUUGUCGGCCGAGCGGGUGAGGGGCGACAAUAAGGAGUACGAGUACUCGGUCGAAUGCCAGGAGGACUCCGCCUGCCCCGCCGCCGAGGAGUCCCUCCCCAUCGAAGUGAUGGUGGACGCCGUCCACAAGCUGAAGUACGAGAACUACACCAGCAGCUUCUUCAUACGGGAUAUCAUCAAGCCCGACCCCCCGAAGAACCUGCAGCUGAAACCCUUGAAGAACUCCAGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGUCCACCCCGCACUCAUACUUCAGCCUGACCUUCUGUGUACAGGUCCAGGGCAAGAGCAAGAGGGAAAAGAAGGAUAGGGUGUUCACCGACAAGACCUCCGCCACGGUGAUCUGUCGGAAAAACGCCAGCAUCUCCGUGCGGGCCCAGGACAGGUACUAUUCCAGCAGCUGGAGCGAGUGGGCCUCCGUCCCCUGCUCCGGCGGCGGUGGCGGGGGCAGCAGGAACCUCCCCGUGGCCACCCCCGAUCCCGGGAUGUUCCCAUGCCUGCACCACAGCCAAAACCUGCUGAGGGCCGUCUCCAAUAUGCUGCAGAAGGCGAGGCAGACCCUGGAGUUCUACCCCUGUACCUCCGAGGAGAUCGACCACGAGGAUAUCACCAAGGACAAGACCUCCACGGUCGAGGCGUGCCUGCCCCUGGAGCUCACGAAGAACGAGAGCUGCCUUAACUCCAGGGAAACCUCGUUUAUCACGAACGGCAGCUGCCUGGCGUCACGGAAGACCUCCUUUAUGAUGGCCCUAUGUCUGUCCUCGAUCUACGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGAUCCCAAGAGGCAGAUUUUCCUGGACCAGAACAUGCUGGCCGUGAUUGACGAGCUGAUGCAGGCGCUGAACUUCAACAGCGAGACAGUGCCGCAGAAGAGCUCCCUGGAGGAGCCGGACUUUUACAAGACCAAGAUAAAGCUGUGCAUCCUGCUCCACGCCUUCAGAAUACGGGCCGUCACCAUCGAUAGGGUGAUGUCUUACCUGAACGCCUCCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 170 hIL12AB_030G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAGCAGCUGGUGAUUAGCUGGUUUAGCCUGGUGUUCCUGGCAAGCCCCCUGGUG T100 tail)GCCAUCUGGGAACUGAAAAAGGACGUGUACGUGGUCGAGCUGGAUUGGUACCCCGACGCCCCCGGCGAAAUGGUGGUGCUGACGUGUGAUACCCCCGAGGAGGACGGGAUCACCUGGACCCUGGAUCAGAGCAGCGAGGUGCUGGGGAGCGGGAAGACCCUGACGAUCCAGGUCAAGGAGUUCGGCGACGCUGGGCAGUACACCUGUCACAAGGGCGGGGAGGUGCUGUCCCACUCCCUGCUGCUCCUGCAUAAGAAAGAGGACGGCAUCUGGUCCACCGACAUCCUCAAGGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGCGGUGUGAGGCGAAGAACUACAGCGGCCGUUUCACCUGCUGGUGGCUGACGACAAUCAGCACCGACUUGACGUUCUCCGUGAAGUCCUCCAGAGGCAGCUCCGACCCCCAAGGGGUGACGUGCGGCGCGGCCACCCUGAGCGCCGAGCGGGUGCGGGGGGACAACAAGGAGUACGAGUACUCCGUGGAGUGCCAGGAGGACAGCGCCUGUCCCGCAGCCGAGGAGUCCCUGCCCAUCGAAGUCAUGGUGGACGCCGUCCACAAGCUGAAGUACGAGAACUACACCAGCAGCUUCUUCAUCCGCGAUAUCAUCAAGCCCGAUCCCCCCAAAAACCUGCAACUGAAGCCGCUGAAGAAUAGCAGGCAGGUGGAGGUGUCCUGGGAGUACCCGGACACCUGGAGCACGCCCCACAGCUAUUUCAGCCUGACCUUUUGCGUGCAGGUCCAGGGGAAGAGCAAGCGGGAGAAGAAGGACCGCGUGUUUACGGACAAAACCAGCGCCACCGUGAUCUGCAGGAAGAACGCCAGCAUCAGCGUGAGGGCCCAGGACAGGUACUACAGCAGCUCCUGGAGCGAGUGGGCCUCCGUGCCCUGUUCCGGAGGCGGCGGGGGCGGUUCCCGGAACCUCCCGGUGGCCACCCCCGACCCGGGCAUGUUCCCGUGCCUGCACCACUCACAGAAUCUGCUGAGGGCCGUGAGCAAUAUGCUGCAGAAGGCAAGGCAGACCCUGGAGUUUUAUCCCUGCACCAGCGAGGAGAUCGACCACGAAGACAUCACCAAGGACAAGACCAGCACAGUGGAGGCCUGCCUGCCCCUGGAACUGACCAAGAACGAGUCCUGUCUGAACUCCCGGGAAACCAGCUUCAUAACCAACGGCUCCUGUCUCGCCAGCAGGAAGACCAGCUUCAUGAUGGCCCUGUGCCUCAGCUCCAUCUACGAGGACCUCAAGAUGUACCAGGUUGAGUUCAAGACCAUGAACGCCAAGCUCCUGAUGGACCCCAAGAGGCAGAUCUUCCUGGACCAGAAUAUGCUGGCCGUGAUCGAUGAGUUAAUGCAGGCGCUGAACUUCAACAGCGAGACGGUGCCCCAAAAGUCCUCGCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUCCUGCACGCCUUCCGAAUCCGGGCCGUAACCAUCGACAGGGUGAUGAGCUAUCUCAACGCCUCCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 171 hIL12AB_031G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAGCAGCUCGUGAUCAGCUGGUUCUCGCUUGUGUUCCUGGCCUCCCCCCUCGUC T100 tail)GCCAUCUGGGAGCUGAAGAAAGACGUGUACGUGGUGGAGCUGGACUGGUAUCCCGACGCCCCGGGGGAGAUGGUGGUGCUGACCUGCGACACCCCGGAAGAGGACGGCAUCACCUGGACGCUCGACCAGUCGUCCGAAGUGCUGGGGUCGGGCAAGACCCUCACCAUCCAGGUGAAGGAGUUCGGAGACGCCGGCCAGUACACCUGUCAUAAGGGGGGGGAGGUGCUGAGCCACAGCCUCCUGCUCCUGCACAAAAAGGAGGACGGCAUCUGGAGCACCGAUAUCCUCAAGGACCAGAAGGAGCCCAAGAACAAGACGUUCCUGAGGUGUGAGGCCAAGAACUACAGCGGGCGGUUCACGUGUUGGUGGCUCACCACCAUCUCCACCGACCUCACCUUCUCCGUGAAGUCAAGCAGGGGCAGCUCCGACCCCCAAGGCGUCACCUGCGGCGCCGCCACCCUGAGCGCCGAGAGGGUCAGGGGGGAUAACAAGGAAUACGAGUACAGUGUGGAGUGCCAAGAGGAUAGCGCCUGUCCCGCCGCCGAAGAGAGCCUGCCCAUCGAAGUGAUGGUGGACGCCGUGCACAAGCUGAAGUACGAGAACUACACCUCCAGCUUCUUCAUCAGGGAUAUCAUCAAGCCCGAUCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCAGGCAGGUGGAGGUGAGCUGGGAGUAUCCCGACACGUGGAGCACCCCGCACAGCUACUUCUCGCUGACCUUCUGCGUGCAGGUGCAAGGGAAGUCCAAGAGGGAGAAGAAGGAUAGGGUGUUCACCGACAAAACGAGCGCCACCGUGAUCUGCCGGAAGAAUGCCAGCAUCUCUGUGAGGGCCCAGGACAGGUACUAUUCCAGCUCCUGGUCGGAGUGGGCCAGCGUGCCCUGUAGCGGCGGGGGCGGGGGCGGCAGCAGGAACCUCCCGGUUGCCACCCCCGACCCCGGCAUGUUUCCGUGCCUGCACCACUCGCAAAACCUGCUGCGCGCGGUCUCCAACAUGCUGCAAAAAGCGCGCCAGACGCUGGAGUUCUACCCCUGCACCAGCGAGGAGAUCGAUCAUGAAGAUAUCACCAAAGACAAGACCUCGACCGUGGAGGCCUGCCUGCCCCUGGAGCUCACCAAGAACGAAAGCUGCCUGAACAGCAGGGAGACAAGCUUCAUCACCAACGGCAGCUGCCUGGCCUCCCGGAAGACCAGCUUCAUGAUGGCCCUGUGCCUGUCCAGCAUCUACGAGGAUCUGAAGAUGUACCAAGUGGAGUUUAAGACCAUGAACGCCAAGCUGUUAAUGGACCCCAAAAGGCAGAUCUUCCUGGAUCAGAACAUGCUGGCCGUCAUCGACGAGCUGAUGCAAGCCCUGAACUUCAACAGCGAGACGGUGCCCCAGAAGAGCAGCCUCGAGGAGCCCGACUUCUAUAAGACCAAGAUAAAGCUGUGCAUUCUGCUGCACGCCUUCAGAAUCAGGGCCGUGACCAUCGAUAGGGUGAUGAGCUACCUGAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 172 hIL12AB_032G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGUC (mRNA withACCAGCAGCUGGUGAUUUCCUGGUUCAGUCUGGUGUUUCUUGCCAGCCCCCUGGUG T100 tail)GCCAUCUGGGAGCUGAAGAAAGACGUAUACGUCGUGGAGCUGGACUGGUAUCCCGACGCUCCCGGCGAGAUGGUGGUCCUCACCUGCGACACCCCAGAGGAGGACGGCAUCACCUGGACCCUGGACCAGAGCUCCGAGGUCCUGGGCAGCGGUAAGACCCUCACCAUCCAGGUGAAGGAGUUUGGUGAUGCCGGGCAGUAUACCUGCCACAAGGGCGGCGAGGUGCUGUCCCACAGCCUCCUGUUACUGCAUAAGAAGGAGGAUGGCAUCUGGAGCACCGACAUCCUCAAGGACCAGAAAGAGCCCAAGAACAAGACCUUUCUGCGGUGCGAGGCGAAAAAUUACUCCGGCCGGUUCACCUGCUGGUGGCUGACCACCAUCAGCACGGACCUGACGUUCUCCGUGAAGUCGAGCAGGGGGAGCUCCGAUCCCCAGGGCGUGACCUGCGGCGCGGCCACCCUGAGCGCCGAGCGCGUCCGCGGGGACAAUAAGGAAUACGAAUAUAGCGUGGAGUGCCAGGAGGACAGCGCCUGCCCCGCGGCCGAGGAGAGCCUCCCGAUCGAGGUGAUGGUGGAUGCCGUCCACAAGCUCAAAUACGAAAACUACACCAGCAGCUUCUUCAUUAGGGACAUCAUCAAGCCCGACCCCCCCAAAAACCUGCAGCUGAAGCCCCUGAAGAACAGCCGCCAGGUCGAGGUGUCAUGGGAGUACCCAGACACCUGGAGCACCCCCCACUCCUACUUCAGCCUGACCUUCUGCGUCCAGGUGCAGGGAAAGUCCAAACGGGAGAAGAAGGAUAGGGUCUUUACCGAUAAGACGUCGGCCACCGUCAUCUGCAGGAAGAACGCCAGCAUAAGCGUGCGGGCGCAGGAUCGGUACUACAGCUCGAGCUGGUCCGAAUGGGCCUCCGUGCCCUGUAGCGGAGGGGGUGGCGGGGGCAGCAGGAACCUGCCCGUGGCCACCCCGGACCCGGGCAUGUUUCCCUGCCUGCAUCACAGUCAGAACCUGCUGAGGGCCGUGAGCAACAUGCUCCAGAAGGCCCGCCAGACCCUGGAGUUUUACCCCUGCACCAGCGAAGAGAUCGAUCACGAAGACAUCACCAAAGACAAGACCUCCACCGUGGAGGCCUGUCUGCCCCUGGAGCUGACCAAGAACGAGAGCUGUCUGAACAGCAGGGAGACCUCCUUCAUCACCAACGGCUCCUGCCUGGCAUCCCGGAAGACCAGCUUCAUGAUGGCCCUGUGUCUGAGCUCUAUCUACGAGGACCUGAAGAUGUACCAGGUCGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGACAGAUAUUCCUGGACCAGAACAUGCUCGCCGUGAUCGAUGAACUGAUGCAAGCCCUGAACUUCAAUAGCGAGACCGUGCCCCAGAAAAGCAGCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAACUGUGCAUACUGCUGCACGCGUUCAGGAUCCGGGCCGUCACCAUCGACCGGGUGAUGUCCUAUCUGAAUGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 173 hIL12AB_033G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAGCAGCUCGUGAUUAGCUGGUUUUCGCUGGUGUUCCUGGCCAGCCCUCUCGUG T100 tail)GCCAUCUGGGAGCUGAAAAAAGACGUGUACGUGGUGGAGCUGGACUGGUACCCGGACGCCCCCGGCGAGAUGGUGGUGCUGACGUGCGACACCCCGGAAGAGGACGGCAUCACCUGGACCCUGGACCAGUCAUCCGAGGUCCUGGGCAGCGGCAAGACGCUCACCAUCCAGGUGAAGGAGUUCGGCGACGCCGGCCAGUACACAUGCCAUAAGGGCGGGGAGGUGCUGAGCCACAGCCUGCUCCUCCUGCACAAGAAGGAGGAUGGCAUCUGGUCUACAGACAUCCUGAAGGACCAGAAAGAGCCCAAGAACAAGACCUUCCUCCGGUGCGAGGCCAAGAACUACUCCGGGCGGUUUACUUGUUGGUGGCUGACCACCAUCAGCACCGACCUCACCUUCAGCGUGAAGAGCUCCCGAGGGAGCUCCGACCCCCAGGGGGUCACCUGCGGCGCCGCCACCCUGAGCGCCGAGCGGGUGAGGGGCGACAACAAGGAGUAUGAAUACAGCGUGGAAUGCCAAGAGGACAGCGCCUGUCCCGCGGCCGAGGAAAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUCCACAAACUCAAGUACGAGAACUACACCAGCAGUUUCUUCAUUCGCGACAUCAUCAAGCCGGACCCCCCCAAAAACCUGCAGCUCAAACCCCUGAAGAACAGCAGGCAGGUGGAGGUCAGCUGGGAGUACCCGGACACCUGGAGCACCCCCCAUAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGCAAGAGCAAACGCGAGAAGAAGGACCGGGUGUUUACCGACAAGACCAGCGCCACGGUGAUCUGCCGAAAGAAUGCAAGCAUCUCCGUGAGGGCGCAGGACCGCUACUACUCUAGCAGCUGGAGCGAGUGGGCCAGCGUGCCCUGCAGCGGUGGCGGCGGAGGCGGCAGCCGUAACCUCCCCGUGGCCACCCCCGACCCCGGCAUGUUCCCGUGUCUGCACCACUCCCAGAACCUGCUGAGGGCCGUCAGCAAUAUGCUGCAGAAGGCCCGGCAGACGCUGGAGUUCUACCCCUGCACCUCCGAGGAGAUCGACCAUGAGGACAUUACCAAGGACAAGACGAGCACUGUGGAGGCCUGCCUGCCCCUGGAGCUCACCAAAAACGAGAGCUGCCUGAAUAGCAGGGAGACGUCCUUCAUCACCAACGGCAGCUGUCUGGCCAGCAGGAAGACCAGCUUCAUGAUGGCCCUGUGCCUCUCCUCCAUAUAUGAGGAUCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGAUCCCAAGAGGCAGAUCUUCCUGGACCAGAAUAUGCUGGCCGUGAUUGACGAGCUGAUGCAGGCCCUGAACUUUAAUAGCGAGACCGUCCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUCUAUAAGACCAAGAUCAAGCUGUGCAUACUGCUGCACGCGUUUAGGAUAAGGGCCGUCACCAUCGACAGGGUGAUGAGCUACCUGAAUGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 174 hIL12AB_034G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAACAGCUGGUGAUCUCCUGGUUCAGCCUGGUGUUCCUCGCCAGCCCCCUGGUG T100 tail)GCCAUCUGGGAGCUGAAGAAAGACGUGUACGUGGUGGAGCUGGACUGGUAUCCCGACGCCCCCGGCGAGAUGGUCGUGCUGACCUGCGACACCCCGGAGGAGGACGGCAUCACCUGGACCCUGGAUCAGUCCUCCGAGGUGCUGGGCAGCGGGAAGACCCUGACCAUCCAGGUGAAAGAGUUCGGAGAUGCCGGCCAGUAUACCUGUCACAAGGGGGGUGAGGUGCUGAGCCAUAGCCUCUUGCUUCUGCACAAGAAGGAGGACGGCAUCUGGUCCACCGACAUCCUCAAGGACCAAAAGGAGCCGAAGAAUAAAACGUUCCUGAGGUGCGAAGCCAAGAACUAUUCCGGACGGUUCACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUCACCUUCUCCGUAAAGUCAAGCAGGGGCAGCUCCGACCCCCAGGGCGUGACCUGCGGAGCCGCCACCCUGAGCGCAGAGAGGGUGAGGGGCGACAACAAGGAGUACGAAUACUCCGUCGAGUGCCAGGAGGACAGCGCCUGCCCCGCCGCCGAGGAAAGUCUGCCCAUCGAGGUGAUGGUGGACGCCGUGCACAAGCUCAAAUACGAGAACUACACCAGCAGCUUCUUCAUCCGGGAUAUCAUCAAGCCCGACCCUCCAAAGAAUCUGCAGCUGAAACCCCUUAAGAACAGCAGGCAGGUGGAGGUCAGCUGGGAGUACCCCGACACCUGGAGCACGCCCCACUCCUACUUUAGCCUGACCUUUUGCGUGCAGGUGCAGGGGAAAAGCAAGCGGGAGAAGAAGGACAGGGUGUUCACCGAUAAGACCUCCGCUACCGUGAUCUGCAGGAAGAACGCCUCAAUCAGCGUGAGGGCCCAGGAUCGGUACUACUCCAGCUCCUGGAGCGAGUGGGCCAGCGUGCCCUGCUCUGGCGGUGGCGGCGGGGGCAGCCGGAACCUGCCGGUGGCCACUCCCGACCCGGGCAUGUUCCCGUGCCUCCACCAUUCCCAGAACCUGCUGCGGGCCGUGUCCAAUAUGCUCCAGAAGGCAAGGCAGACCCUGGAGUUCUACCCCUGCACCAGCGAGGAGAUCGAUCACGAGGACAUCACCAAAGACAAAACCAGCACGGUCGAGGCCUGCCUGCCCCUGGAACUCACCAAGAACGAAAGCUGUCUCAACAGCCGCGAGACCAGCUUCAUAACCAACGGUUCCUGUCUGGCCUCCCGCAAGACCAGCUUUAUGAUGGCCCUCUGUCUGAGCUCCAUCUAUGAAGACCUGAAAAUGUACCAGGUGGAGUUCAAAACCAUGAACGCCAAGCUUCUGAUGGACCCCAAGAGGCAGAUCUUCCUGGAUCAGAACAUGCUGGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUUAACUCCGAGACCGUGCCCCAGAAAAGCAGCCUGGAAGAGCCCGAUUUCUACAAAACGAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCCGGAUCCGUGCGGUGACCAUCGAUAGGGUGAUGAGCUACCUGAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 175 hIL12AB_035G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAACAGCUGGUAAUCAGCUGGUUCAGCCUGGUUUUCCUCGCGUCGCCCCUGGUG T100 tail)GCCAUCUGGGAGUUAAAGAAGGACGUGUACGUGGUGGAGCUGGAUUGGUACCCCGACGCCCCGGGCGAGAUGGUCGUGCUCACCUGCGAUACCCCCGAGGAGGACGGGAUCACCUGGACCCUGGACCAAUCCAGCGAGGUGCUGGGCAGCGGCAAGACCCUGACCAUACAGGUGAAGGAAUUUGGGGACGCCGGGCAGUACACCUGCCACAAGGGCGGGGAAGUGCUGUCCCACUCCCUCCUGCUGCUGCAUAAGAAGGAGGACGGCAUCUGGAGCACCGACAUCCUGAAGGACCAAAAGGAGCCCAAGAACAAGACCUUCCUGAGGUGCGAGGCCAAAAACUAUUCCGGCCGCUUUACCUGUUGGUGGCUGACCACCAUCUCCACCGAUCUGACCUUCAGCGUGAAGUCGUCUAGGGGCUCCUCCGACCCCCAGGGCGUAACCUGCGGCGCCGCGACCCUGAGCGCCGAGAGGGUGCGGGGCGAUAACAAAGAGUACGAGUACUCGGUGGAGUGCCAGGAGGACAGCGCCUGUCCGGCGGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUCCACAAGCUGAAGUACGAGAACUACACCAGUUCGUUCUUCAUCAGGGACAUCAUCAAGCCGGACCCCCCCAAGAACCUCCAGCUGAAGCCCCUGAAGAACAGCAGGCAGGUGGAAGUGUCCUGGGAGUAUCCCGACACCUGGAGCACCCCCCACAGCUACUUCAGCCUGACCUUUUGCGUGCAGGUGCAGGGCAAAAGCAAGAGGGAAAAGAAGGACCGGGUGUUCACCGAUAAGACGAGCGCCACCGUUAUCUGCAGGAAGAACGCCUCCAUAAGCGUGAGGGCGCAGGACCGUUACUACAGCAGCAGCUGGAGUGAGUGGGCAAGCGUGCCCUGUAGCGGCGGGGGCGGGGGCGGGUCCCGCAACCUCCCCGUCGCCACCCCCGACCCAGGCAUGUUUCCGUGCCUGCACCACAGCCAGAACCUGCUGCGGGCCGUUAGCAACAUGCUGCAGAAGGCCAGGCAGACCCUCGAGUUCUAUCCCU GCACAUCUGAGGAGAUCGAGCACGAAGACAUCACUAAGGAUAAGACCUCCACCGUGGAGGCCUGUCUGCCCCUCGAGCUGACCAAGAAUGAAUCCUGCCUGAACAGCCGAGAGACCAGCUUUAUCACCAACGGCUCCUGCCUGGCCAGCAGGAAGACCUCCUUCAUGAUGGCCCUGUGCCUCUCCAGCAUCUACGAGGAUCUGAAGAUGUACCAGGUAGAGUUCAAGACGAUGAACGCCAAGCUCCUGAUGGACCCCAAGAGGCAGAUAUUCCUGGACCAGAACAUGCUGGCGGUGAUCGACGAGCUGAUGCAGGCCCUGAAUUUCAACAGCGAGACGGUGCCACAGAAGUCCAGCCUGGAGGAGCCAGACUUCUACAAGACCAAGAUCAAACUGUGCAUCCUCCUGCACGCGUUCAGGAUCCGCGCCGUCACCAUAGACAGGGUGAUGAGUUAUCUGAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 176 hIL12AB_036G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withAUCAGCAGCUGGUAAUCAGCUGGUUUAGCCUGGUGUUCCUGGCCAGCCCACUGGUG T100 tail)GCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAACUGGACUGGUACCCCGACGCCCCUGGCGAGAUGGUGGUACUGACCUGUGACACCCCGGAGGAAGACGGUAUCACCUGGACCCUGGAUCAGAGCUCCGAGGUGCUGGGCUCCGGCAAGACACUGACCAUCCAAGUUAAGGAAUUUGGGGACGCCGGCCAGUACACCUGCCACAAGGGGGGCGAGGUGCUGUCCCACUCCCUGCUGCUUCUGCAUAAGAAGGAGGAUGGCAUCUGGUCCACCGACAUACUGAAGGACCAGAAGGAGCCCAAGAAUAAGACCUUCCUGAGAUGCGAGGCCAAGAACUACUCGGGAAGGUUCACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUGACCUUCUCCGUGAAGAGCUCCCGGGGCAGCUCCGACCCCCAGGGCGUAACCUGUGGGGCCGCUACCCUGUCCGCCGAGAGGGUCCGGGGCGACAACAAGGAAUACGAGUACAGCGUGGAGUGCCAGGAGGACUCCGCCUGCCCCGCCGCCGAGGAGUCGCUGCCCAUAGAGGUGAUGGUGGACGCCGUGCACAAGCUCAAGUACGAGAAUUACACCAGCAGCUUCUUUAUCAGGGACAUAAUUAAGCCGGACCCCCCAAAGAAUCUGCAGCUGAAGCCCCUGAAGAAUAGCCGGCAGGUGGAAGUGUCCUGGGAGUACCCCGACACCUGGAGCACCCCCCACUCCUAUUUCUCACUGACAUUCUGCGUGCAGGUGCAAGGGAAAAGCAAGAGGGAGAAGAAGGAUAGGGUGUUCACCGACAAGACAAGCGCCACCGUGAUCUGCCGAAAAAAUGCCAGCAUCAGCGUGAGGGCCCAGGAUCGGUAUUACAGCAGCUCCUGGAGCGAGUGGGCCAGCGUGCCCUGUUCCGGCGGGGGAGGGGGCGGCUCCCGGAACCUGCCGGUGGCCACCCCCGACCCUGGCAUGUUCCCCUGCCUGCAUCACAGCCAGAACCUGCUCCGGGCCGUGUCGAACAUGCUGCAGAAGGCCCGGCAGACCCUCGAGUUUUACCCCUGCACCAGCGAAGAGAUCGACCACGAAGACAUAACCAAGGACAAGACCAGCACGGUGGAGGCCUGCCUGCCCCUGGAGCUUACCAAAAACGAGUCCUGCCUGAACAGCCGGGAAACCAGCUUCAUAACGAACGGGAGCUGCCUGGCCUCCAGGAAGACCAGCUUCAUGAUGGCGCUGUGUCUGUCCAGCAUAUACGAGGAUCUGAAGAUGUAUCAGGUGGAAUUCAAAACUAUGAAUGCCAAGCUCCUGAUGGACCCCAAGAGGCAGAUCUUCCUGGACCAGAACAUGCUAGCCGUGAUCGACGAGCUGAUGCAGGCCCUCAACUUCAACUCGGAGACGGUGCCCCAGAAGUCCAGCCUCGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUACUGCUGCAUGCCUUCAGGAUAAGGGCGGUGACUAUCGACAGGGUCAUGUCCUACCUGAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 177 hIL12AB_037G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAACAACUGGUGAUCAGCUGGUUCUCCCUGGUGUUCCUGGCCAGCCCCCUGGUG T100 tail)GCCAUCUGGGAGCUCAAAAAAGACGUGUACGUGGUGGAGCUCGAUUGGUACCCAGACGCGCCGGGGGAAAUGGUGGUGCUGACCUGCGACACCCCAGAGGAGGAUGGCAUCACGUGGACGCUGGAUCAGUCCAGCGAGGUGCUGGGGAGCGGCAAGACGCUCACCAUCCAGGUGAAGGAAUUUGGCGACGCGGGCCAGUAUACCUGUCACAAGGGCGGCGAGGUGCUGAGCCACUCCCUGCUGCUGCUGCACAAGAAGGAGGAUGGGAUCUGGUCAACCGAUAUCCUGAAAGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGCGCUGCGAGGCCAAGAACUAUAGCGGCAGGUUCACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUGACCUUCAGCGUGAAAUCCUCCAGGGGCAGCAGCGACCCCCAGGGCGUGACCUGCGGUGCCGCCACGCUCUCCGCCGAGCGAGUGAGGGGUGACAACAAGGAGUACGAGUACAGCGUGGAAUGUCAGGAGGACAGCGCCUGUCCCGCCGCCGAGGAGUCGCUGCCCAUCGAGGUGAUGGUCGACGCGGUGCACAAGCUCAAAUACGAGAAUUACACCAGCAGCUUCUUCAUCAGGGACAUCAUCAAGCCCGACCCCCCCAAGAACCUGCAGCUGAAGCCCUUGAAGAACAGCAGGCAGGUGGAGGUGAGCUGGGAGUACCCGGACACCUGGAGCACCCCCCACUCCUACUUCAGCCUGACGUUCUGUGUGCAGGUGCAGGGGAAGUCCAAGAGGGAGAAGAAGGACCGGGUGUUCACCGACAAGACCAGCGCCACCGUGAUAUGCCGCAAGAACGCGUCCAUCAGCGUUCGCGCCCAGGACCGCUACUACAGCAGCUCCUGGUCCGAAUGGGCCAGCGUGCCCUGCAGCGGUGGAGGGGGCGGGGGCUCCAGGAAUCUGCCGGUGGCCACCCCCGACCCCGGGAUGUUCCCGUGUCUGCAUCACUCCCAGAACCUGCUGCGGGCCGUGAGCAAUAUGCUGCAGAAGGCCAGGCAGACGCUCGAGUUCUACCCCUGCACCUCCGAAGAGAUCGACCAUGAGGACAUCACCAAGGACAAGACCAGCACCGUGGAGGCCUGCCUCCCCCUGGAGCUGACCAAAAACGAGAGCUGCCUGAACUCCAGGGAGACCAGCUUUAUAACCAACGGCAGCUGCCUCGCCUCCAGGAAGACCUCGUUUAUGAUGGCCCUCUGCCUGUCCAGCAUCUACGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCGAAGUUGCUCAUGGACCCCAAGAGGCAGAUCUUCCUGGACCAGAACAUGCUCGCGGUGAUCGACGAGCUGAUGCAAGCCCUGAACUUCAACAGCGAGACCGUGCCCCAGAAGAGCAGCCUGGAAGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCCGGAUCCGGGCCGUGACCAUCGACAGGGUGAUGAGCUACCUCAACGCCUCCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 178 hIL12AB_038G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAGCAGCUCGUGAUCAGCUGGUUCUCCCUCGUCUUCCUGGCCUCCCCGCUGGUG T100 tail)GCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGCUGGACUGGUAUCCCGACGCCCCCGGCGAGAUGGUGGUGCUGACGUGCGACACACCAGAAGAGGACGGGAUCACAUGGACCCUGGAUCAGUCGUCCGAGGUGCUGGGGAGCGGCAAGACCCUCACCAUCCAAGUGAAGGAGUUCGGGGACGCCGGCCAGUACACCUGCCACAAGGGCGGGGAGGUGCUCUCCCAUAGCCUGCUCCUCCUGCACAAAAAGGAGGAUGGCAUCUGGAGCACCGACAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACAUUUCUCAGGUGUGAGGCCAAGAACUAUUCGGGCAGGUUUACCUGUUGGUGGCUCACCACCAUCUCUACCGACCUGACGUUCUCCGUCAAGUCAAGCAGGGGGAGCUCGGACCCCCAGGGGGUGACAUGUGGGGCCGCCACCCUGAGCGCGGAGCGUGUCCGCGGCGACAACAAGGAGUACGAGUAUUCCGUGGAGUGCCAGGAGGACAGCGCCUGCCCCGCCGCCGAGGAGUCCCUGCCCAUAGAGGUGAUGGUGGACGCCGUCCACAAGUUGAAGUACGAAAAUUAUACCUCCUCGUUCUUCAUUAGGGACAUCAUCAAGCCUGACCCCCCGAAGAACCUACAACUCAAGCCCCUCAAGAACUCCCGCCAGGUGGAGGUGUCCUGGGAGUACCCCGACACCUGGUCCACCCCGCACAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUCCAGGGGAAGAGCAAGCGUGAAAAGAAAGACAGGGUGUUCACCGACAAGACGAGCGCCACCGUGAUCUGCAGGAAAAACGCCUCCAUCUCCGUGCGCGCCCAGGACAGGUACUACAGUAGCUCCUGGAGCGAAUGGGCCAGCGUGCCGUGCAGCGGCGGGGGAGGAGGCGGCAGUCGCAACCUGCCCGUGGCCACCCCCGACCCCGGCAUGUUCCCAUGCCUGCACCACAGCCAGAACCUGCUGAGGGCAGUCAGCAAUAUGCUGCAGAAGGCCAGGCAGACCCUGGAGUUUUAUCCCUGCACCAGCGAGGAGAUCGACCACGAGGACAUCACCAAGGACAAGACCUCCACCGUCGAGGCCUGCCUGCCACUGGAGCUGACCAAAAACGAGAGCUGCCUGAACUCCAGGGAGACCUCCUUCAUCACCAACGGGAGCUGCCUGGCCAGCCGGAAGACCAGCUUCAUGAUGGCGCUGUGCCUCAGCAGCAUCUACGAGGAUCUCAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCGAAGCUGCUGAUGGACCCCAAGCGGCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUGAUUGACGAGCUCAUGCAGGCCCUGAACUUCAAUAGCGAGACCGUCCCCCAAAAGAGCAGCCUGGAGGAACCCGACUUCUACAAAACGAAGAUCAAGCUCUGCAUCCUGCUGCACGCCUUCCGGAUCCGGGCCGUGACCAUCGAUCGUGUGAUGAGCUACCUGAACGCCUCGUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 179 hIL12AB_039G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withACCAGCAGCUCGUCAUCUCCUGGUUUAGCCUGGUGUUUCUGGCCUCCCCCCUGGUC T100 tail)GCCAUCUGGGAGCUGAAGAAAGACGUGUACGUGGUGGAGCUGGACUGGUACCCGGACGCUCCCGGGGAGAUGGUGGUGCUGACCUGCGACACCCCCGAGGAGGACGGCAUCACCUGGACCCUGGACCAGAGCUCCGAGGUGCUGGGGAGCGGCAAGACCCUGACCAUUCAGGUGAAAGAGUUCGGCGACGCCGGCCAAUAUACCUGCCACAAGGGGGGGGAGGUCCUGUCGCAUUCCCUGCUGCUGCUUCACAAAAAGGAGGAUGGCAUCUGGAGCACCGACAUCCUGAAGGACCAGAAAGAACCCAAGAACAAGACGUUCCUGCGCUGCGAGGCCAAGAACUACAGCGGCCGGUUCACCUGUUGGUGGCUGACCACCAUCUCCACCGACCUGACUUUCUCGGUGAAGAGCAGCCGCGGGAGCAGCGACCCCCAGGGAGUGACCUGCGGCGCCGCCACCCUGAGCGCCGAAAGGGUGAGGGGCGACAAUAAAGAGUACGAGUAUUCCGUGGAGUGCCAGGAGGACAGCGCCUGUCCCGCCGCCGAGGAGUCCCUGCCUAUCGAGGUGAUGGUCGACGCGGUGCACAAGCUCAAGUACGAAAACUACACCAGCAGCUUUUUCAUCAGGGAUAUCAUCAAACCAGACCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAAAACAGCAGGCAGGUGGAAGUGAGCUGGGAAUACCCCGAUACCUGGUCCACCCCCCACAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGGAAGUCCAAGCGGGAGAAGAAAGAUCGGGUGUUCACGGACAAGACCAGCGCCACCGUGAUUUGCAGGAAAAACGCCAGCAUCUCCGUGAGGGCUCAGGACAGGUACUACAGCUCCAGCUGGAGCGAGUGGGCCUCCGUGCCUUGCAGCGGGGGAGGAGGCGGCGGCAGCAGGAAUCUGCCCGUCGCAACCCCCGACCCCGGCAUGUUCCCCUGCCUGCACCACAGCCAGAAUCUGCUGCGAGCCGUGAGCAACAUGCUCCAGAAGGCCCGGCAGACGCUGGAGUUCUACCCCUGCACCUCCGAGGAGAUCGACCACGAGGACAUCACCAAGGAUAAGACGAGCACCGUCGAGGCCUGUCUCCCCCUGGAGCUCACCAAGAACGAGUCCUGCCUGAAUAGCAGGGAGACGUCCUUCAUAACCAACGGCAGCUGUCUGGCGUCCAGGAAGACCAGCUUCAUGAUGGCCCUCUGCCUGAGCUCCAUCUACGAGGACCUCAAGAUGUACCAGGUCGAGUUCAAGACCAUGAACGCAAAACUGCUCAUGGAUCCAAAGAGGCAGAUCUUUCUGGACCAGAACAUGCUGGCCGUGAUCGAUGAACUCAUGCAGGCCCUGAAUUUCAAUUCCGAGACCGUGCCCCAGAAGAGCUCCCUGGAGGAACCCGACUUCUACAAAACAAAGAUCAAGCUGUGUAUCCUCCUGCACGCCUUCCGGAUCAGGGCCGUCACCAUUGACCGGGUGAUGUCCUACCUGAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 180 hIL12AB_040G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCC (mRNA withAUCAGCAGCUGGUGAUCAGCUGGUUCAGCCUCGUGUUCCUCGCCAGCCCCCUCGUG T100 tail)GCCAUCUGGGAGCUGAAAAAGGACGUGUACGUGGUGGAGCUGGACUGGUAUCCCGACGCCCCGGGCGAGAUGGUGGUGCUGACCUGCGACACCCCCGAGGAGGACGGCAUUACCUGGACACUGGACCAGAGCAGCGAGGUCCUGGGCAGCGGGAAGACCCUGACAAUUCAGGUGAAGGAGUUCGGCGACGCCGGACAGUACACGUGCCACAAGGGGGGGGAGGUGCUGUCCCACAGCCUCCUCCUGCUGCACAAGAAGGAGGAUGGCAUCUGGAGCACCGACAUCCUGAAGGAUCAGAAGGAGCCCAAGAACAAGACCUUUCUGAGAUGCGAGGCCAAGAAUUACAGCGGCCGUUUCACCUGCUGGUGGCUCACCACCAUCAGCACCGACCUGACCUUCAGCGUGAAAUCCUCCAGGGGCUCCUCCGACCCGCAGGGAGUGACCUGCGGCGCCGCCACACUGAGCGCCGAGCGGGUCAGAGGGGACAACAAGGAGUACGAGUACAGCGUUGAGUGCCAGGAGGACAGCGCCUGUCCCGCGGCCGAGGAAUCCCUGCCCAUCGAGGUGAUGGUGGACGCAGUGCACAAGCUGAAGUACGAGAACUAUACCUCGAGCUUCUUCAUCCGGGAUAUCAUUAAGCCCGAUCCCCCGAAGAACCUGCAGCUCAAACCCCUGAAGAACAGCAGGCAGGUGGAGGUCUCCUGGGAGUACCCCGACACAUGGUCCACCCCCCAUUCCUAUUUCUCCCUGACCUUUUGCGUGCAGGUGCAGGGCAAGAGCAAGAGGGAGAAAAAGGACAGGGUGUUCACCGACAAGACCUCCGCCACCGUGAUCUGCCGUAAGAACGCUAGCAUCAGCGUCAGGGCCCAGGACAGGUACUAUAGCAGCUCCUGGUCCGAGUGGGCCAGCGUCCCGUGCAGCGGCGGGGGCGGUGGAGGCUCCCGGAACCUCCCCGUGGCCACCCCGGACCCCGGGAUGUUUCCCUGCCUGCAUCACAGCCAGAACCUGCUGAGGGCCGUGUCCAACAUGCUGCAGAAGGCCAGGCAGACACUCGAGUUUUACCCCUGCACCAGCGAGGAGAUCGACCACGAAGACAUCACCAAGGACAAGACCUCCACCGUGGAGGCAUGCCUGCCCCUGGAGCUGACCAAAAACGAAAGCUGUCUGAACUCCAGGGAGACCUCCUUUAUCACGAACGGCUCAUGCCUGGCCUCCAGAAAGACCAGCUUCAUGAUGGCCCUGUGCCUGAGCUCCAUCUACGAGGACUUGAAAAUGUACCAGGUCGAGUUCAAGACCAUGAACGCCAAGCUGCUCAUGGACCCCAAAAGGCAGAUCUUUCUGGACCAGAAUAUGCUGGCCGUGAUCGACGAGCUCAUGCAAGCCCUGAAUUUCAACAGCGAGACCGUGCCCCAGAAGUCCUCCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUACUCCUGCACGCGUUUAGGAUCAGGGCGGUGACCAUCGAUAGGGUGAUGAGCUACCUGAAUGCCUCCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG G* = a 5′terminal guanosine cap 181 mIL12ABATGTGTCCTCAGAAGCTAACCATCTCCTGGTTTGCCATCGTTTTGTTGGTGTCTCC 80TM-ACTCATGGCCATGTGGGAGCTCGAGAAAGACGTTTACGTTGTAGAGGTGGACTGGA nolinker_CTCCCGACGCCCCGGGCGAAACAGTGAACCTCACCTGTGACACGCCTGAAGAAGAT V5_GACATCACCTGGACCTCAGACCAGAGACATGGAGTCATAGGCTCTGGAAAGACCCT NucleotideGACCATCACTGTCAAAGAGTTCCTAGATGCTGGCCAGTACACCTGCCACAAAGGAG SequenceGCGAGACTCTGAGCCACTCACATCTGCTGCTCCACAAGAAGGAGAATGGAATTTGGTCCACTGAGATCCTGAAGAACTTCAAGAATAAGACTTTCCTGAAGTGTGAAGCACCAAATTACTCCGGACGGTTCACGTGCTCATGGCTGGTGCAAAGAAACATGGACTTGAAGTTCAACATCAAGAGCAGTAGCAGTTCCCCTGACTCTCGGGCAGTGACATGTGGAATGGCGTCTCTGTCTGCAGAGAAGGTCACACTGGACCAAAGGGACTATGAGAAGTATTCAGTGTCCTGCCAGGAGGATGTCACCTGCCCAACTGCCGAGGAGACACTGCCCATTGAACTGGCGTTGGAAGCACGGCAGCAGAATAAATATGAGAACTACAGCACCAGCTTCTTCATCAGGGACATCATCAAACCAGACCCGCCCAAGAACTTGCAGATGAAGCCTTTGAAGAACTCACAGGTGGAGGTCAGCTGGGAGTACCCGGACTCCTGGAGCACTCCCCATTCCTACTTCTCCCTCAAGTTCTTTGTTCGAATCCAGCGCAAGAAAGAGAAGATGAAGGAGACAGAGGAGGGGTGTAACCAGAAAGGTGCGTTCCTCGTAGAGAAGACATCTACCGAAGTCCAATGCAAAGGCGGGAATGTCTGCGTGCAAGCTCAGGATCGCTATTACAATTCCTCATGCAGCAAGTGGGCATGTGTTCCCTGCAGGGTCCGATCCGGAGGCGGAGGGAGCGGTGGTGGAGGCAGCGGAGGAGGTGGATCAAGGGTCATTCCAGTCTCTGGACCAGCTAGATGTCTTAGCCAGTCCCGAAACCTGCTGAAGACCACGGACGACATGGTGAAGACGGCCAGAGAGAAACTGAAACATTATTCCTGCACCGCAGAGGATATCGATCACGAAGATATCACACGGGACCAAACCAGCACATTGAAGACCTGTTTACCACTGGAACTACACAAGAACGAGAGTTGCCTGGCTACTAGAGAGACTTCTTCCACAACAAGAGGGAGCTGCCTGCCACCACAGAAGACGTCTTTGATGATGACCCTGTGCCTTGGTAGCATCTATGAGGACCTCAAGATGTACCAGACAGAGTTCCAGGCCATCAACGCAGCACTTCAGAATCACAACCATCAGCAGATCATTTTAGACAAGGGCATGCTGGTGGCCATCGATGAGCTGATGCAATCATTGAATCATAACGGTGAGACATTGCGCCAGAAACCTCCTGTGGGAGAAGCAGACCCTTACAGAGTGAAGATGAAGCTCTGCATCCTGCTTCACGCCTTCAGCACCCGCGTCGTCACTATCAACAGGGTGATGGGCTATCTGAGCTCCGCCACCCTGGTGCTGTTCGGCGCCGGCTTCGGTGCAGTGATCACCGTGGTGGTGATCGTCGTCATCATCGGGAAACCAATTCCAAATCCCCTCCTGGGGTTGGATAGCACC 182 mIL12AB-MCPQKLTISWFAIVLLVSPLMA MWELEKDVYVVEVDWTPDAPGETVNLTCDTPEED 80TM-DITWTSDQRHGVIGSGKTLTITVKEFLDAGQYTCHKGGETLSHSHLLLHKKENGIW nolinker_V5STEILKNFKNKTFLKCEAPNYSGRFTCSWLVQRNMDLKFNIKSSSSSPDSRAVTCG Amino acidMASLSAEKVTLDQRDYEKYSVSCQEDVTCPTAEETLPIELALEARQQNKYENYSTS SequenceFFIRDIIKPDPPKNLQMKPLKNSQVEVSWEYPDSWSTPHSYFSLKFFVRIQRKKEKMKETEEGCNQKGAFLVEKTSTEVQCKGGNVCVQAQDRYYNSSCSKWACVPCRVRS

IDHEDITRDQTSTLKTCLPLELHKNESCLATRETSSTTRGSCLPPQKTSLMMTLCLGSIYEDLKMYQTEFQAINAALQNHNHQQIILDKGMLVAIDELMQSLNHNGETLRQK

Italic: signal peptide; Underline: IL12B; Dashed underlineand Italic: linker; Bold: IL12A; Double underline:Transmembrane domain; Bold and Italic: V5 tag 183 mIL12AB-ATGTGTCCTCAGAAGCTAACCATCTCCTGGTTTGCCATCGTTTTGCTTGTGTCTCC PTM_v5ACTCATGGCCATGTGGGAGCTCGAGAAAGACGTTTACGTTGTAGAGGTGGACTGGA miR122CTCCCGACGCCCCAGGAGAAACAGTGAACCTCACCTGTGACACGCCTGAAGAAGAT NucleotideGACATCACCTGGACCTCAGACCAGAGACATGGAGTCATAGGCTCTGGAAAGACCCT SequenceGACCATCACTGTCAAAGAGTTCCTAGATGCTGGCCAGTACACCTGCCACAAAGGAGGCGAGACTCTGAGCCACTCACATCTGCTGCTCCACAAGAAGGAGAATGGAATTTGGTCCACCGAAATCCTGAAGAACTTCAAGAATAAGACTTTCCTGAAGTGTGAAGCACCAAATTACTCCGGACGGTTCACGTGCTCATGGCTGGTGCAAAGAAACATGGACTTGAAGTTCAACATCAAGAGCAGTAGCAGTTCCCCTGACTCTCGGGCAGTGACATGTGGAATGGCGTCTCTGTCTGCAGAGAAGGTCACACTGGACCAAAGGGACTATGAGAAGTATTCAGTGTCCTGCCAGGAGGATGTCACCTGCCCAACTGCCGAGGAAACTCTGCCCATTGAACTGGCGTTGGAAGCACGGCAGCAGAATAAATATGAGAACTACAGCACCAGCTTCTTCATCAGGGACATCATCAAACCAGACCCGCCCAAGAACTTGCAGATGAAGCCTTTGAAGAACTCACAGGTGGAGGTCAGCTGGGAGTACCCAGACTCCTGGAGCACTCCCCATTCCTACTTCTCCCTCAAGTTCTTTGTTCGAATCCAGCGCAAGAAAGAGAAGATGAAGGAGACAGAGGAGGGGTGTAACCAGAAAGGTGCGTTCCTCGTAGAGAAGACATCTACCGAAGTCCAATGCAAAGGCGGGAATGTCTGCGTGCAAGCTCAGGATCGCTATTACAATTCCTCATGCAGCAAGTGGGCATGTGTTCCCTGCAGGGTCCGATCCGGAGGCGGAGGGTCTGGAGGAGGAGGTTCTGGAGGTGGTGGCAGTAGGGTCATTCCAGTCTCTGGACCTGCAAGGTGTCTTAGCCAGTCCCGAAACCTGCTGAAGACCACAGACGATATGGTGAAGACGGCCAGAGAGAAACTGAAACATTATTCCTGCACAGCAGAGGACATCGATCATGAAGATATTACACGGGACCAAACCAGCACATTGAAGACCTGTTTACCACTGGAACTACACAAGAACGAGAGTTGCCTGGCTACTAGAGAGACTTCTTCCACAACAAGAGGGAGCTGCCTGCCACCACAGAAGACGTCTTTGATGATGACCCTGTGCCTTGGTAGCATCTACGAGGATCTCAAGATGTACCAGACAGAGTTCCAGGCCATCAACGCAGCACTTCAGAATCACAACCATCAGCAGATCATTTTAGACAAGGGCATGCTGGTGGCCATCGATGAGCTGATGCAATCTCTGAATCATAATGGCGAGACACTTCGCCAGAAACCTCCTGTGGGAGAAGCAGACCCTTACAGAGTGAAGATGAAGCTCTGCATCCTGCTTCACGCCTTCAGCACCCGCGTCGTCACTATTAACAGGGTGATGGGCTATCTGAGCTCCGCCTCTGGTGGCGGATCAGGCGGCGGCGGCTCTGGCGGCGGTGGAAGCGGAGGTGGCGGGTCTGGCGGAGGTTCACTGCAGGTAGTAGTGATCAGCGCCATCCTGGCCCTGGTGGTGCTGACCGTGATCTCATTGATCATCTTGATTATGCTGTGGGGCGGAGGAGGCAGCGGGAAACCAATTCCAAATCCCCTCCTGGGGTTGGATAGCACC 184 mIL12AB-MCPQKLTISWFAIVLLVSPLMA MWELEKDVYVVEVDWTPDAPGETVNLTCDTPEED PTM_v5DITWTSDQRHGVIGSGKTLTITVKEFLDAGQYTCHKGGETLSHSHLLLHKKENGIW miR122STEILKNFKNKTFLKCEAPNYSGRFTCSWLVQRNMDLKFNIKSSSSSPDSRAVTCG Amino AcidMASLSAEKVTLDQRDYEKYSVSCQEDVTCPTAEETLPIELALEARQQNKYENYSTS SequenceFFIRDIIKPDPPKNLQMKPLKNSQVEVSWEYPDSWSTPHSYFSLKFFVRIQRKKEKMKETEEGCNQKGAFLVEKTSTEVQCKGGNVCVQAQDRYYNSSCSKWACVPCRVRS

IDHEDITRDQTSTLKTCLPLELHKNESCLATRETSSTTRGSCLPPQKTSLMMTLCLGSIYEDLKMYQTEFQAINAALQNHNHQQIILDKGMLVAIDELMQSLNHNGETLRQK

Italic: signal peptide; Underline: IL12B; Dashed underlineand Italic: linker; Bold: IL12A; Double underline:Transmembrane domain; Bold and Italic: V5 tag 185 mIL12AB-ATGTGTCCTCAGAAGCTAACCATCTCCTGGTTTGCCATCGTTTTGTTAGTGTCTCC 8TM_v5_miRACTCATGGCCATGTGGGAGCTCGAGAAAGACGTTTACGTTGTAGAGGTGGACTGGA 122CTCCCGACGCCCCAGGCGAAACAGTGAACCTCACCTGTGACACGCCTGAAGAAGAT NucleotideGACATCACCTGGACCTCAGACCAGAGACATGGAGTCATAGGCTCTGGAAAGACCCT sequenceGACCATCACTGTCAAAGAGTTCCTAGATGCTGGCCAGTACACCTGCCACAAAGGAGGCGAGACTCTGAGCCACTCACATCTGCTGCTCCACAAGAAGGAGAATGGAATTTGGTCCACAGAAATTTTAAAGAACTTCAAGAACAAGACTTTCCTGAAGTGTGAAGCACCAAATTACTCCGGACGGTTCACGTGCTCATGGCTGGTGCAAAGAAACATGGACTTGAAGTTCAACATCAAGAGCAGTAGCAGTTCCCCTGACTCTCGGGCAGTGACATGTGGAATGGCGTCTCTGTCTGCAGAGAAGGTCACACTGGACCAAAGGGACTATGAGAAGTATTCAGTGTCCTGCCAGGAGGATGTCACCTGCCCAACTGCCGAGGAGACTCTGCCCATTGAACTGGCGTTGGAAGCACGGCAGCAGAATAAATATGAGAACTACAGCACCAGCTTCTTCATCAGGGACATCATCAAACCAGACCCGCCCAAGAACTTGCAGATGAAGCCTTTGAAGAACTCACAGGTGGAGGTCAGCTGGGAGTACCCAGACTCCTGGAGCACTCCCCATTCCTACTTCTCCCTCAAGTTCTTTGTTCGAATCCAGCGCAAGAAAGAGAAGATGAAGGAGACAGAGGAGGGGTGTAACCAGAAAGGTGCGTTCCTCGTAGAGAAGACATCTACCGAAGTCCAATGCAAAGGCGGGAATGTCTGCGTGCAAGCTCAGGATCGCTATTACAATTCCTCATGCAGCAAGTGGGCATGTGTTCCCTGCAGGGTCCGATCCGGAGGCGGAGGGAGTGGAGGAGGTGGCTCTGGCGGCGGTGGAAGTAGGGTCATTCCAGTCTCTGGACCTGCACGCTGTCTTAGCCAGTCCCGAAACCTGCTGAAGACCACAGACGACATGGTGAAGACGGCCAGAGAGAAACTGAAACATTATTCCTGCACAGCGGAAGACATAGATCACGAGGATATCACACGGGACCAAACCAGCACATTGAAGACCTGTTTACCACTGGAACTACACAAGAACGAGAGTTGCCTGGCTACTAGAGAGACTTCTTCCACAACAAGAGGGAGCTGCCTGCCACCACAGAAGACGTCTTTGATGATGACCCTGTGCCTTGGTAGCATCTATGAGGATCTGAAGATGTACCAGACAGAGTTCCAGGCCATCAACGCAGCACTTCAGAATCACAACCATCAGCAGATCATTTTAGACAAGGGCATGCTGGTGGCCATCGATGAGCTGATGCAGTCCCTGAATCATAATGGTGAAACGTTGCGCCAGAAACCTCCTGTGGGAGAAGCAGACCCTTACAGAGTGAAGATGAAGCTCTGCATCCTGCTTCACGCCTTCAGCACCCGCGTCGTGACTATAAACAGGGTGATGGGCTATCTGAGCTCCGCCTCTGGTGGCGGATCAGGAGGAGGTGGATCCGGTGGCGGTGGTTCCGGAGGTGGTGGATCGGGTGGTGGCTCACTGCAGATCTACATCTGGGCCCCGCTGGCCGGCATCTGCGTGGCCCTGCTGCTGAGCCTGATCATCACCCTGATCTGCTACGGTGGAGGCGGTAGCGGGAAACCAATTCCAAATCCCCTCCTGGGGTTGGATAGCACC 186 mIL12AB-MCPQKLTISWFAIVLLVSPLMA MWELEKDVYVVEVDWTPDAPGETVNLTCDTPEED 8TM_v5_miRDITWTSDQRHGVIGSGKTLTITVKEFLDAGQYTCHKGGETLSHSHLLLHKKENGIW 122STEILKNFKNKTFLKCEAPNYSGRFTCSWLVQRNMDLKFNIKSSSSSPDSRAVTCG Amino AcidMASLSAEKVTLDQRDYEKYSVSCQEDVTCPTAEETLPIELALEARQQNKYENYSTS sequenceFFIRDIIKPDPPKNLQMKPLKNSQVEVSWEYPDSWSTPHSYFSLKFFVRIQRKKEKMKETEEGCNQKGAFLVEKTSTEVQCKGGNVCVQAQDRYYNSSCSKWACVPCRVRS

IDHEDITRDQTSTLKTCLPLELHKNESCLATRETSSTTRGSCLPPQKTSLMMTLCLGSIYEDLKMYQTEFQAINAALQNHNHQQIILDKGMLVAIDELMQSLNHNGETLRQK

Italic: signal peptide; Underline: IL12B; Dashed underlineand Italic: linker; Bold: IL12A; Double underline:Transmembrane domain; Bold and Italic: V5 tag 187 Xba1 TCTAGArestriction  site 188 EcoRI GAATTC 189 EcoRII CCWGG (W = A or T) 190HindIII AAGCTT 191 T7 RNA GnnnnWnCRnCTCnCnnWnD polymerase(n = any nucleotide; R = A or G; W = A or T; D = A or G or T but not C)192 Linker (amino GGGGG acid sequence) 193 Gly/ser linker(G_(n)S)_(m) n = 1-100; m = 1-100 (amino acid sequence) 194Gly/ser linker (GGGGS)_(o) o = 1-5 (amino acid sequence) 195Gly/ser linker GGSGGGGSGG (amino acid sequence) 196 Gly/ser linkerGGSGGGGG (amino acid sequence) 197 Gly/ser linker GSGSGSGS (amino acidsequence) 198 Gly-rich linker (Gly)_(p) p = 1-100 (amino acid sequence)199 Linker (amino (EAAAK)_(q) q = 1-100 acid sequence) 200 Linker (aminoGGGGSLVPRGSGGGGS acid sequence) 201 Linker (amino GSGSGS acid sequence)202 Linker (amino GGGGSLVPRGSGGGG acid sequence) 203 Linker (aminoGGSGGHMGSGG acid sequence) 204 Linker (amino GGSGGSGGSGG acid sequence)205 Linker (amino GGSGG acid sequence) 206 Linker (amino GSGSGSGSacid sequence) 207 Linker (amino GGGSEGGGSEGGGSEGGG acid sequence) 208Linker (amino AAGAATAA acid sequence) 209 Linker (amino GGSSGacid sequence) 210 Linker (amino GSGGGTGGGSG acid sequence) 211Linker (amino GSGSGSGSGGSG acid sequence) 212 Linker (aminoGSGGSGSGGSGGSG acid sequence) 213 Linker (amino GSGGSGGSGGSGGSacid sequence) 214 Linker (amino GGGGGGS acid sequence) 215Polynucleotide ATCCCG 216 Kozak CCR(A/G)CCAUGG R = purine consensussequence 217 Linker (amino GGGGGG acid sequence) 218 Linker (aminoGGGGGGG acid sequence) 219 Linker (amino GGGGGGGG acid sequence) 220hIL12AB_041 ATGTGCCACCAGCAGCTGGTGATCAGCTGGTTCAGCCTGGTGTTCCTGGCCAGCCC ORFCCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGTTGGATTGGTACCCCGACGCCCCCGGCGAGATGGTGGTGCTGACCTGCGACACCCCCGAGGAGGACGGCATCACCTGGACCCTGGACCAGAGCAGCGAGGTGCTGGGCAGCGGCAAGACCCTGACCATCCAGGTGAAGGAGTTCGGCGACGCCGGCCAGTACACCTGCCACAAGGGCGGCGAGGTGCTGAGCCACAGCCTGCTGCTGCTGCACAAGAAGGAGGACGGCATCTGGAGCACCGACATCCTGAAGGACCAGAAGGAGCCCAAGAACAAGACCTTCCTGAGATGCGAGGCCAAGAACTACAGCGGCAGATTCACCTGCTGGTGGCTGACCACCATCAGCACCGACCTGACCTTCAGCGTGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGTGACCTGCGGCGCCGCCACCCTGAGCGCCGAGAGAGTGAGAGGCGACAACAAGGAGTACGAGTACAGCGTGGAGTGCCAGGAAGATAGCGCCTGCCCCGCCGCCGAGGAGAGCCTGCCCATCGAGGTGATGGTGGACGCCGTGCACAAGCTGAAGTACGAGAACTACACCAGCAGCTTCTTCATCAGAGATATCATCAAGCCCGACCCCCCCAAGAACCTGCAGCTGAAGCCCCTGAAGAACAGCCGGCAGGTGGAGGTGAGCTGGGAGTACCCCGACACCTGGAGCACCCCCCACAGCTACTTCAGCCTGACCTTCTGCGTGCAGGTGCAGGGCAAGAGCAAGAGAGAGAAGAAAGATAGAGTGTTCACCGACAAGACCAGCGCCACCGTGATCTGCAGAAAGAACGCCAGCATCAGCGTGAGAGCCCAAGATAGATACTACAGCAGCAGCTGGAGCGAGTGGGCCAGCGTGCCCTGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCTGCCCGTGGCCACCCCCGACCCCGGCATGTTCCCCTGCCTGCACCACAGCCAGAACCTGCTGAGAGCCGTGAGCAACATGCTGCAGAAGGCCCGGCAGACCCTGGAGTTCTACCCCTGCACCAGCGAGGAGATCGACCACGAAGATATCACCAAAGATAAGACCAGCACCGTGGAGGCCTGCCTGCCCCTGGAGCTGACCAAGAACGAGAGCTGCCTGAACAGCAGAGAGACCAGCTTCATCACCAACGGCAGCTGCCTGGCCAGCAGAAAGACCAGCTTCATGATGGCCCTGTGCCTGAGCAGCATCTACGAGGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAACGCCAAGCTGCTGATGGACCCCAAGCGGCAGATCTTCCTGGACCAGAACATGCTGGCCGTGATCGACGAGCTGATGCAGGCCCTGAACTTCAACAGCGAGACCGTGCCCCAGAAGAGCAGCCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATCCTGCTGCACGCCTTCAGAATCAGAGCCGTGACCATCGACAGAGTGATGAGCTACCTGAACGCCAGC 221 hIL12AB_041AUGUGCCACCAGCAGCUGGUGAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCAGCCC mRNA ORFCCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGUUGGAUUGGUACCCCGACGCCCCCGGCGAGAUGGUGGUGCUGACCUGCGACACCCCCGAGGAGGACGGCAUCACCUGGACCCUGGACCAGAGCAGCGAGGUGCUGGGCAGCGGCAAGACCCUGACCAUCCAGGUGAAGGAGUUCGGCGACGCCGGCCAGUACACCUGCCACAAGGGCGGCGAGGUGCUGAGCCACAGCCUGCUGCUGCUGCACAAGAAGGAGGACGGCAUCUGGAGCACCGACAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGAGAUGCGAGGCCAAGAACUACAGCGGCAGAUUCACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUGACCUUCAGCGUGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGUGACCUGCGGCGCCGCCACCCUGAGCGCCGAGAGAGUGAGAGGCGACAACAAGGAGUACGAGUACAGCGUGGAGUGCCAGGAAGAUAGCGCCUGCCCCGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUGCACAAGCUGAAGUACGAGAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAGCCCGACCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCCGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGAGCACCCCCCACAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGCAAGAGCAAGAGAGAGAAGAAAGAUAGAGUGUUCACCGACAAGACCAGCGCCACCGUGAUCUGCAGAAAGAACGCCAGCAUCAGCGUGAGAGCCCAAGAUAGAUACUACAGCAGCAGCUGGAGCGAGUGGGCCAGCGUGCCCUGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCUGCCCGUGGCCACCCCCGACCCCGGCAUGUUCCCCUGCCUGCACCACAGCCAGAACCUGCUGAGAGCCGUGAGCAACAUGCUGCAGAAGGCCCGGCAGACCCUGGAGUUCUACCCCUGCACCAGCGAGGAGAUCGACCACGAAGAUAUCACCAAAGAUAAGACCAGCACCGUGGAGGCCUGCCUGCCCCUGGAGCUGACCAAGAACGAGAGCUGCCUGAACAGCAGAGAGACCAGCUUCAUCACCAACGGCAGCUGCCUGGCCAGCAGAAAGACCAGCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGGCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAGACCGUGCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCAGAAUCAGAGCCGUGACCAUCGACAGAGUGAUGAGCUACCUGAACGCCAGC 222 3UTR-018 +UAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCC miR-122-5p

binding site CUUUGAAUAAAGUCUGAGUGGGCGGC 223 3UTR-018 +UAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCC miR-122-3p

binding site CUUUGAAUAAAGUCUGAGUGGGCGGC 224 3UTR-019 +UGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCA miR-122GCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGU binding siteGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 225 Human CD80 TYCFAPRCRERRRNERLRRESVRPVIntracellular Domain 226 HumanQKKPRYEIRWKVIESVSSDGHEYIYVDPMQLPYDSTWELPRDQLVLGRTLGSGAFG PGFRBQVVEATAHGLSHSQATMKVAVKMLKSTARSSEKQALMSELKIMSHLGPHLNVVNLL IntracellularGACTKGGPIYIITEYCRYGDLVDYLHRNKHTFLQHHSDKRRPPSAELYSNALPVGL Domain (WT)PLPSHVSLTGESDGGYMDMSKDESVDYVPMLDMKGDVKYADIESSNYMAPYDNYVPSAPERTCRATLINESPVLSYMDLVGFSYQVANGMEFLASKNCVHRDLAARNVLICEGKLVKICDFGLARDIMRDSNYISKGSTFLPLKWMAPESIFNSLYTTLSDVWSFGILLWEIFTLGGTPYPELPMNEQFYNAIKRGYRMAQPAHASDEIYEIMQKCWEEKFEIRPPFSQLVLLLERLLGEGYKKKYQQVDEEFLRSDHPAILRSQARLPGFHGLRSPLDTSSVLYTAVQPNEGDNDYIIPLPDPKPEVADEGPLEGSPSLASSTLNEVNTSSTISCDSPLEPQDEPEPEPQLELQVEPEPELEQLPDSGCPAPRAEAEDSFL 227 HumanQKKPRYEIRWKVIESVSSDGHE PGFRB Intracellular Domain (E570tr) 228 HumanQKKPRYEIRWKVIESVSSDGHEFIFVDPMQLPYDSTWELPRDQLVLGRTLGSGAFG PGFRBQVVEATAHGLSHSQATMKVAVAMLKSTARSSEKQALMSELKIMSHLGPHLNVVNLL IntracellularGACTKGGPIYIITEYCRYGDLVDYLHRNKHTFLQHHSDKRRPPSAELYSNALPVGL DomainPLPSHVSLTGESDGG (G739tr) 229 Linker SGGGSGGGGSGGGGSGGGGSGGGSLQ 230V5 tag GKPIPNPLLGLDST 231 G4S Linker GGGGS 232 Murine CD8IYIWAPLAGICVALLLSLIITLI Transmembrane Domain 233 MurineVVVISAILALWLTVISLIILIMLW PDGFR Transmembrane domain 234 Murine CD80TLVLFGAGFGAVITVVVIVVII Transmembrane domain 235 Murine CD80KCFCKHRSCFRRNEASRETNNSLTFGPEEALA intracellular domain 236 mIL12AB-ATGTGTCCTCAGAAGCTAACCATCTCCTGGTTTGCCATCGTTTTGTTGGTGTCTCC 80TM-ICDACTCATGGCCATGTGGGAGCTCGAGAAAGACGTTTACGTTGTAGAGGTGGACTGGA NucleotideCTCCCGACGCCCCGGGCGAAACAGTGAACCTCACCTGTGACACGCCTGAAGAAGAT SequenceGACATCACCTGGACCTCAGACCAGAGACATGGAGTCATAGGCTCTGGAAAGACCCTGACCATCACTGTCAAAGAGTTCCTAGATGCTGGCCAGTACACCTGCCACAAAGGAGGCGAGACTCTGAGCCACTCACATCTGCTGCTCCACAAGAAGGAGAATGGAATTTGGTCCACTGAGATCCTGAAGAACTTCAAGAATAAGACTTTCCTGAAGTGTGAAGCACCAAATTACTCCGGACGGTTCACGTGCTCATGGCTGGTGCAAAGAAACATGGACTTGAAGTTCAACATCAAGAGCAGTAGCAGTTCCCCTGACTCTCGGGCAGTGACATGTGGAATGGCGTCTCTGTCTGCAGAGAAGGTCACACTGGACCAAAGGGACTATGAGAAGTATTCAGTGTCCTGCCAGGAGGATGTCACCTGCCCAACTGCCGAGGAGACACTGCCCATTGAACTGGCGTTGGAAGCACGGCAGCAGAATAAATATGAGAACTACAGCACCAGCTTCTTCATCAGGGACATCATCAAACCAGACCCGCCCAAGAACTTGCAGATGAAGCCTTTGAAGAACTCACAGGTGGAGGTCAGCTGGGAGTACCCGGACTCCTGGAGCACTCCCCATTCCTACTTCTCCCTCAAGTTCTTTGTTCGAATCCAGCGCAAGAAAGAGAAGATGAAGGAGACAGAGGAGGGGTGTAACCAGAAAGGTGCGTTCCTCGTAGAGAAGACATCTACCGAAGTCCAATGCAAAGGCGGGAATGTCTGCGTGCAAGCTCAGGATCGCTATTACAATTCCTCATGCAGCAAGTGGGCATGTGTTCCCTGCAGGGTCCGATCCGGAGGCGGAGGGAGCGGTGGTGGAGGCAGCGGAGGAGGTGGATCAAGGGTCATTCCAGTCTCTGGACCAGCTAGATGTCTTAGCCAGTCCCGAAACCTGCTGAAGACCACGGACGACATGGTGAAGACGGCCAGAGAGAAACTGAAACATTATTCCTGCACCGCAGAGGATATCGATCACGAAGATATCACACGGGACCAAACCAGCACATTGAAGACCTGTTTACCACTGGAACTACACAAGAACGAGAGTTGCCTGGCTACTAGAGAGACTTCTTCCACAACAAGAGGGAGCTGCCTGCCACCACAGAAGACGTCTTTGATGATGACCCTGTGCCTTGGTAGCATCTATGAGGACCTCAAGATGTACCAGACAGAGTTCCAGGCCATCAACGCAGCACTTCAGAATCACAACCATCAGCAGATCATTTTAGACAAGGGCATGCTGGTGGCCATCGATGAGCTGATGCAATCATTGAATCATAACGGTGAGACATTGCGCCAGAAACCTCCTGTGGGAGAAGCAGACCCTTACAGAGTGAAGATGAAGCTCTGCATCCTGCTTCACGCCTTCAGCACCCGCGTCGTCACTATCAACAGGGTGATGGGCTATCTGAGCTCCGCCAGCGGTGGCGGAAGCGGTGGAGGCGGCAGCGGCGGTGGTGGTAGCGGCGGCGGCGGCTCCGGCGGAGGGAGCCTGCAGACCCTGGTGCTGTTCGGCGCCGGCTTCGGTGCAGTGATCACCGTGGTGGTGATCGTCGTCATCATCAAGTGCTTCTGCAAGCACAGAAGCTGCTTCAGAAGAAACGAGGCCAGCAGAGAAACCAACAACAGCCTAACATTCGGCCCAGAAGAGGCTCTGGCC 237 mIL12AB- MCPQKLTISWFAIVLLVSPLMAMWELEKDVYVVEVDWTPDAPGETVNLTCDTPEED 80TM-ICDDITWTSDQRHGVIGSGKTLTITVKEFLDAGQYTCHKGGETLSHSHLLLHKKENGIW Amino AcidSTEILKNFKNKTFLKCEAPNYSGRFTCSWLVQRNMDLKFNIKSSSSSPDSRAVTCG SequenceMASLSAEKVTLDQRDYEKYSVSCQEDVTCPTAEETLPIELALEARQQNKYENYSTSFFIRDIIKPDPPKNLQMKPLKNSQVEVSWEYPDSWSTPHSYFSLKFFVRIQRKKEKMKETEEGCNQKGAFLVEKTSTEVQCKGGNVCVQAQDRYYNSSCSKWACVPCRVRS

IDHEDITRDQTSTLKTCLPLELHKNESCLATRETSSTTRGSCLPPQKTSLMMTLCLGSIYEDLKMYQTEFQAINAALQNHNHQQIILDKGMLVAIDELMQSLNHNGETLRQK

Italic: signal peptide; Underline: IL12B; Dashed underlineand Italic: linker; Bold: IL12A; Double underline:Transmembrane domain; Bold and Italic: Intracellular domain 238 IgK-ATGGAGACTGACACCCTGCTGCTGTGGGTGCTGTTACTTTGGGTTCCCGGCAGCAC mscIL12AB-CGGCTACCCCTACGACGTGCCCGACTACGCCATGTGGGAGCTCGAGAAAGACGTTT 80TM-ICDACGTTGTAGAGGTGGACTGGACTCCCGACGCCCCGGGCGAAACAGTGAACCTCACC NucleotideTGTGACACGCCTGAAGAAGATGACATCACCTGGACCTCAGACCAGAGACATGGAGT SequenceCATAGGCTCTGGAAAGACCCTGACCATCACTGTCAAAGAGTTCCTAGATGCTGGCCAGTACACCTGCCACAAAGGAGGCGAGACTCTGAGCCACTCACATCTGCTGCTCCACAAGAAGGAGAATGGAATTTGGTCCACTGAGATCCTGAAGAACTTCAAGAATAAGACTTTCCTGAAGTGTGAAGCACCAAATTACTCCGGACGGTTCACGTGCTCATGGCTGGTGCAAAGAAACATGGACTTGAAGTTCAACATCAAGAGCAGTAGCAGTTCCCCTGACTCTCGGGCAGTGACATGTGGAATGGCGTCTCTGTCTGCAGAGAAGGTCACACTGGACCAAAGGGACTATGAGAAGTATTCAGTGTCCTGCCAGGAGGATGTCACCTGCCCAACTGCCGAGGAGACACTGCCCATTGAACTGGCGTTGGAAGCACGGCAGCAGAATAAATATGAGAACTACAGCACCAGCTTCTTCATCAGGGACATCATCAAACCAGACCCGCCCAAGAACTTGCAGATGAAGCCTTTGAAGAACTCACAGGTGGAGGTCAGCTGGGAGTACCCGGACTCCTGGAGCACTCCCCATTCCTACTTCTCCCTCAAGTTCTTTGTTCGAATCCAGCGCAAGAAAGAGAAGATGAAGGAGACAGAGGAGGGGTGTAACCAGAAAGGTGCGTTCCTCGTAGAGAAGACATCTACCGAAGTCCAATGCAAAGGCGGGAATGTCTGCGTGCAAGCTCAGGATCGCTATTACAATTCCTCATGCAGCAAGTGGGCATGTGTTCCCTGCAGGGTCCGAGGAAGCACCAGCGGTTCCGGCAAACCAGGTAGCGGAGAGGGCAGCACCAAGGGCAGGGTCATTCCAGTCTCTGGACCAGCTAGATGTCTTAGCCAGTCCCGAAACCTGCTGAAGACCACGGACGACATGGTGAAGACGGCCAGAGAGAAACTGAAACATTATTCCTGCACCGCAGAGGATATCGATCACGAAGATATCACACGGGACCAAACCAGCACATTGAAGACCTGTTTACCACTGGAACTACACAAGAACGAGAGTTGCCTGGCTACTAGAGAGACTTCTTCCACAACAAGAGGGAGCTGCCTGCCACCACAGAAGACGTCTTTGATGATGACCCTGTGCCTTGGTAGCATCTATGAGGACCTCAAGATGTACCAGACAGAGTTCCAGGCCATCAACGCAGCACTTCAGAATCACAACCATCAGCAGATCATTTTAGACAAGGGCATGCTGGTGGCCATCGATGAGCTGATGCAATCATTGAATCATAACGGTGAGACATTGCGCCAGAAACCTCCTGTGGGAGAAGCAGACCCTTACAGAGTGAAGATGAAGCTCTGCATCCTGCTTCACGCCTTCAGCACCCGCGTCGTCACTATCAACAGGGTGATGGGCTATCTGAGCTCCGCCACCCTGGTGCTGTTCGGCGCCGGCTTCGGTGCAGTGATCACCGTGGTGGTGATCGTCGTCATCATCAAGTGCTTTTGCAAGCACAGAAGCTGTTTCAGAAGAAACGAGGCCAGCAGAGAAACCAACAACTCCCTGACTTTCGGGCCCGAGGAAGCCCTCGCC 239 IgK- METDTLLLWVLLLWVPGSTG

MWELEKDVYVVEVDWTPDAPGETVNLT mscIL12AB-CDTPEEDDITWTSDQRHGVIGSGKTLTITVKEFLDAGQYTCHKGGETLSHSHLLLH 80TM-ICDKKENGIWSTEILKNFKNKTFLKCEAPNYSGRFTCSWLVQRNMDLKFNIKSSSSSPD Amino AcidSRAVTCGMASLSAEKVTLDQRDYEKYSVSCQEDVTCPTAEETLPIELALEARQQNK SequenceYENYSTSFFIRDIIKPDPPKNLQMKPLKNSQVEVSWEYPDSWSTPHSYFSLKFFVRIQRKKEKMKETEEGCNQKGAFLVEKTSTEVQCKGGNVCVQAQDRYYNSSCSKWACV

KHYSCTAEDIDHEDITRDQTSTLKTCLPLELHKNESCLATRETSSTTRGSCLPPQKTSLMMTLCLGSIYEDLKMYQTEFQAINAALQNHNHQQIILDKGMLVAIDELMQSLN

Italic: signal peptide; Underline: IL12B; Dashed underlineand Italic: linker; Bold: IL12A; Double underline:Transmembrane domain; Bold and Italic: Intracellulardomain; Bold underline: epitope tag 240 hIL12AB-ATGTGCCACCAGCAGCTGGTGATCAGCTGGTTCAGCCTGGTGTTCCTGGCCAGCCC 8TMCCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGTTGGATTGGT NucleotideACCCCGACGCCCCCGGCGAGATGGTGGTGCTGACCTGCGACACCCCCGAGGAGGAC SequenceGGCATCACCTGGACCCTGGACCAGAGCAGCGAGGTGCTGGGCAGCGGCAAGACCCTGACCATCCAGGTGAAGGAGTTCGGCGACGCCGGCCAGTACACCTGCCACAAGGGCGGCGAGGTGCTGAGCCACAGCCTGCTGCTGCTGCACAAGAAGGAGGACGGCATCTGGAGCACCGACATCCTGAAGGACCAGAAGGAGCCCAAGAACAAGACCTTCCTGAGATGCGAGGCCAAGAACTACAGCGGCAGATTCACCTGCTGGTGGCTGACCACCATCAGCACCGACCTGACCTTCAGCGTGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGTGACCTGCGGCGCCGCCACCCTGAGCGCCGAGAGAGTGAGAGGCGACAACAAGGAGTACGAGTACAGCGTGGAGTGCCAGGAAGATAGCGCCTGCCCCGCCGCCGAGGAGAGCCTGCCCATCGAGGTGATGGTGGACGCCGTGCACAAGCTGAAGTACGAGAACTACACCAGCAGCTTCTTCATCAGAGATATCATCAAGCCCGACCCCCCCAAGAACCTGCAGCTGAAGCCCCTGAAGAACAGCCGGCAGGTGGAGGTGAGCTGGGAGTACCCCGACACCTGGAGCACCCCCCACAGCTACTTCAGCCTGACCTTCTGCGTGCAGGTGCAGGGCAAGAGCAAGAGAGAGAAGAAAGATAGAGTGTTCACCGACAAGACCAGCGCCACCGTGATCTGCAGAAAGAACGCCAGCATCAGCGTGAGAGCCCAAGATAGATACTACAGCAGCAGCTGGAGCGAGTGGGCCAGCGTGCCCTGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCTGCCCGTGGCCACCCCCGACCCCGGCATGTTCCCCTGCCTGCACCACAGCCAGAACCTGCTGAGAGCCGTGAGCAACATGCTGCAGAAGGCCCGGCAGACCCTGGAGTTCTACCCCTGCACCAGCGAGGAGATCGACCACGAAGATATCACCAAAGATAAGACCAGCACCGTGGAGGCCTGCCTGCCCCTGGAGCTGACCAAGAACGAGAGCTGCCTGAACAGCAGAGAGACCAGCTTCATCACCAACGGCAGCTGCCTGGCCAGCAGAAAGACCAGCTTCATGATGGCCCTGTGCCTGAGCAGCATCTACGAGGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAACGCCAAGCTGCTGATGGACCCCAAGCGGCAGATCTTCCTGGACCAGAACATGCTGGCCGTGATCGACGAGCTGATGCAGGCCCTGAACTTCAACAGCGAGACCGTGCCCCAGAAGAGCAGCCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATCCTGCTGCACGCCTTCAGAATCAGAGCCGTGACCATCGACAGAGTGATGAGCTACCTGAACGCCAGCTCTGGTGGCGGATCAGGCGGCGGCGGTTCAGGAGGCGGTGGAAGTGGAGGTGGCGGGTCTGGCGGAGGTTCACTGCAGATCTACATCTGGGCTCCACTGGCCGGCACCTGCGGCGTGCTGCTGCTGAGCCTGGTGATCACCCTGTACTGCTACGGGAAACCAATTCCAAATCCCCTCCTGGGGTTGGATAGCACC 241 hIL12AB-MCHQQLVISWFSLVELASPLVA IWELKKDVYVVELDWYPDAPGEMVVLTCDTPEED 8TM AminoGITWTLDQSSEVLGSGKTLTIQVKEFGDAGQYTCHKGGEVLSHSLLLLHKKEDGIW Acid SequenceSTDILKDQKEPKNKTFLRCEAKNYSGRFTCWWLTTISTDLTFSVKSSRGSSDPQGVTCGAATLSAERVRGDNKEYEYSVECQEDSACPAAEESLPIEVMVDAVHKLKYENYTSSFFIRDIIKPDPPKNLQLKPLKNSRQVEVSWEYPDTWSTPHSYFSLTFCVQVQGK

NLPVATPDPGMFPCLHHSQNLLRAVSNMLQKARQTLEFYPCTSEEIDHEDITKDKTSTVEACLPLELTKNESCLNSRETSFITNGSCLASRKTSFMMALCLSSIYEDLKMYQVEFKTMNAKLLMDPKRQIFLDQNMLAVIDELMQALNFNSETVPQKSSLEEPDFYKT

Italic: signal peptide; Underline: IL12B; Dashed underlineand Italic: linker; Bold: IL12A; Double underline:Transmembrane domain; Bold and Italic: Epitope tag 242 hIL12AB-ATGTGCCACCAGCAGCTGGTGATCAGCTGGTTCAGCCTGGTGTTCCTGGCCAGCCC 8TM noCCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGTTGGATTGGT epitope tagACCCCGACGCCCCCGGCGAGATGGTGGTGCTGACCTGCGACACCCCCGAGGAGGAC NucleotideGGCATCACCTGGACCCTGGACCAGAGCAGCGAGGTGCTGGGCAGCGGCAAGACCCT SequenceGACCATCCAGGTGAAGGAGTTCGGCGACGCCGGCCAGTACACCTGCCACAAGGGCGGCGAGGTGCTGAGCCACAGCCTGCTGCTGCTGCACAAGAAGGAGGACGGCATCTGGAGCACCGACATCCTGAAGGACCAGAAGGAGCCCAAGAACAAGACCTTCCTGAGATGCGAGGCCAAGAACTACAGCGGCAGATTCACCTGCTGGTGGCTGACCACCATCAGCACCGACCTGACCTTCAGCGTGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGTGACCTGCGGCGCCGCCACCCTGAGCGCCGAGAGAGTGAGAGGCGACAACAAGGAGTACGAGTACAGCGTGGAGTGCCAGGAAGATAGCGCCTGCCCCGCCGCCGAGGAGAGCCTGCCCATCGAGGTGATGGTGGACGCCGTGCACAAGCTGAAGTACGAGAACTACACCAGCAGCTTCTTCATCAGAGATATCATCAAGCCCGACCCCCCCAAGAACCTGCAGCTGAAGCCCCTGAAGAACAGCCGGCAGGTGGAGGTGAGCTGGGAGTACCCCGACACCTGGAGCACCCCCCACAGCTACTTCAGCCTGACCTTCTGCGTGCAGGTGCAGGGCAAGAGCAAGAGAGAGAAGAAAGATAGAGTGTTCACCGACAAGACCAGCGCCACCGTGATCTGCAGAAAGAACGCCAGCATCAGCGTGAGAGCCCAAGATAGATACTACAGCAGCAGCTGGAGCGAGTGGGCCAGCGTGCCCTGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCTGCCCGTGGCCACCCCCGACCCCGGCATGTTCCCCTGCCTGCACCACAGCCAGAACCTGCTGAGAGCCGTGAGCAACATGCTGCAGAAGGCCCGGCAGACCCTGGAGTTCTACCCCTGCACCAGCGAGGAGATCGACCACGAAGATATCACCAAAGATAAGACCAGCACCGTGGAGGCCTGCCTGCCCCTGGAGCTGACCAAGAACGAGAGCTGCCTGAACAGCAGAGAGACCAGCTTCATCACCAACGGCAGCTGCCTGGCCAGCAGAAAGACCAGCTTCATGATGGCCCTGTGCCTGAGCAGCATCTACGAGGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAACGCCAAGCTGCTGATGGACCCCAAGCGGCAGATCTTCCTGGACCAGAACATGCTGGCCGTGATCGACGAGCTGATGCAGGCCCTGAACTTCAACAGCGAGACCGTGCCCCAGAAGAGCAGCCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATCCTGCTGCACGCCTTCAGAATCAGAGCCGTGACCATCGACAGAGTGATGAGCTACCTGAACGCCAGCTCTGGTGGCGGATCAGGCGGCGGCGGTTCAGGAGGCGGTGGAAGTGGAGGTGGCGGGTCTGGCGGAGGTTCACTGCAGATCTACATCTGGGCTCCACTGGCCGGCACCTGCGGCGTGCTGCTGCTGAGCCTGGTGATCAC CCTGTACTGCTAC243 hIL12AB- MCHQQLVISWFSLVFLASPLVA IWELKKDVYVVELDWYPDAPGEMVVLTCDTPEED8TM no GITWTLDQSSEVLGSGKTLTIQVKEFGDAGQYTCHKGGEVLSHSLLLLHKKEDGIWepitope tag STDILKDQKEPKNKTFLRCEAKNYSGRFTCWWLTTISTDLTFSVKSSRGSSDPQGVAmino Acid TCGAATLSAERVRGDNKEYEYSVECQEDSACPAAEESLPIEVMVDAVHKLKYENYTSequence SSFFIRDIIKPDPPKNLQLKPLKNSRQVEVSWEYPDTWSTPHSYFSLTFCVQVQGK

NLPVATPDPGMFPCLHHSQNLLRAVSNMLQKARQTLEFYPCTSEEIDHEDITKDKTSTVEACLPLELTKNESCLNSRETSFITNGSCLASRKTSFMMALCLSSIYEDLKMYQVEFKTMNAKLLMDPKRQIFLDQNMLAVIDELMQALNFNSETVPQKSSLEEPDFYKT

Italic: signal peptide; Underline: IL12B; Dashed underlineand Italic: linker; Bold: IL12A; Double underline: Transmembrane domain244 h12AB-80TID ATGTGCCACCAGCAGCTGGTGATCAGCTGGTTCAGCCTGGTGTTCCTGGCCAGCCCNucleotide CCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGTTGGATTGGTSequence 1 ACCCCGACGCCCCCGGCGAGATGGTGGTGCTGACCTGCGACACCCCCGAGGAGGACmatched GGCATCACCTGGACCCTGGACCAGAGCAGCGAGGTGCTGGGCAGCGGCAAGACCCTGACCATCCAGGTGAAGGAGTTCGGCGACGCCGGCCAGTACACCTGCCACAAGGGCGGCGAGGTGCTGAGCCACAGCCTGCTGCTGCTGCACAAGAAGGAGGACGGCATCTGGAGCACCGACATCCTGAAGGACCAGAAGGAGCCCAAGAACAAGACCTTCCTGAGATGCGAGGCCAAGAACTACAGCGGCAGATTCACCTGCTGGTGGCTGACCACCATCAGCACCGACCTGACCTTCAGCGTGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGTGACCTGCGGCGCCGCCACCCTGAGCGCCGAGAGAGTGAGAGGCGACAACAAGGAGTACGAGTACAGCGTGGAGTGCCAGGAAGATAGCGCCTGCCCCGCCGCCGAGGAGAGCCTGCCCATCGAGGTGATGGTGGACGCCGTGCACAAGCTGAAGTACGAGAACTACACCAGCAGCTTCTTCATCAGAGATATCATCAAGCCCGACCCCCCCAAGAACCTGCAGCTGAAGCCCCTGAAGAACAGCCGGCAGGTGGAGGTGAGCTGGGAGTACCCCGACACCTGGAGCACCCCCCACAGCTACTTCAGCCTGACCTTCTGCGTGCAGGTGCAGGGCAAGAGCAAGAGAGAGAAGAAAGATAGAGTGTTCACCGACAAGACCAGCGCCACCGTGATCTGCAGAAAGAACGCCAGCATCAGCGTGAGAGCCCAAGATAGATACTACAGCAGCAGCTGGAGCGAGTGGGCCAGCGTGCCCTGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCTGCCCGTGGCCACCCCCGACCCCGGCATGTTCCCCTGCCTGCACCACAGCCAGAACCTGCTGAGAGCCGTGAGCAACATGCTGCAGAAGGCCCGGCAGACCCTGGAGTTCTACCCCTGCACCAGCGAGGAGATCGACCACGAAGATATCACCAAAGATAAGACCAGCACCGTGGAGGCCTGCCTGCCCCTGGAGCTGACCAAGAACGAGAGCTGCCTGAACAGCAGAGAGACCAGCTTCATCACCAACGGCAGCTGCCTGGCCAGCAGAAAGACCAGCTTCATGATGGCCCTGTGCCTGAGCAGCATCTACGAGGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAACGCCAAGCTGCTGATGGACCCCAAGCGGCAGATCTTCCTGGACCAGAACATGCTGGCCGTGATCGACGAGCTGATGCAGGCCCTGAACTTCAACAGCGAGACCGTGCCCCAGAAGAGCAGCCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATCCTGCTGCACGCCTTCAGAATCAGAGCCGTGACCATCGACAGAGTGATGAGCTACCTGAACGCCAGCTCTGGTGGCGGATCAGGCGGCGGCGGTTCAGGAGGCGGTGGAAGTGGAGGTGGCGGGTCTGGCGGAGGTTCACTGCAGCTGCTGCCCAGCTGGGCCATCACCCTGATCAGCGTGAACGGCATCTTCGTGATCTGCTGCCTGACCTACTGCTTCGCCCCTCGATGCAGAGAGAGAAGAAGAAACGAGAGACTGAGAAGAGAGAGCGTGCGACCCGTG 245 h12AB-80TIDATGTGCCACCAGCAGCTGGTGATCAGCTGGTTCAGCCTGGTGTTCCTGGCCAGCCC NucleotideTCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGCTGGACTGGT Sequence 2ACCCTGACGCCCCTGGCGAGATGGTGGTGCTGACCTGCGACACCCCTGAGGAGGAC SE_IL12_041GGCATCACCTGGACCCTGGACCAGAGCAGCGAGGTGCTGGGCAGCGGCAAGACCCTGACCATCCAGGTGAAGGAGTTCGGCGACGCCGGCCAGTACACCTGCCACAAGGGCGGAGAGGTGTTAAGCCACAGCCTGCTCTTGCTACACAAGAAGGAGGACGGTATTTGGAGCACCGACATCCTGAAGGACCAGAAGGAGCCTAAGAACAAGACCTTCCTGAGGTGCGAGGCCAAGAACTACAGCGGCAGATTCACCTGCTGGTGGCTGACCACCATCTCCACGGACCTGACCTTCAGCGTTAAGAGTAGCAGAGGCAGCAGCGACCCTCAGGGCGTGACTTGTGGCGCCGCCACCCTGAGCGCCGAGAGAGTGAGAGGCGACAACAAGGAGTACGAGTACAGCGTGGAGTGCCAGGAGGACAGCGCCTGCCCTGCCGCCGAGGAGAGCCTGCCTATCGAGGTGATGGTGGACGCCGTGCACAAGCTGAAGTACGAGAATTACACCTCATCCTTCTTCATCAGAGACATCATCAAGCCTGACCCTCCAAAGAATCTGCAGCTGAAGCCTCTGAAGAACAGCAGACAGGTGGAGGTGAGCTGGGAGTATCCGGATACCTGGAGCACACCTCACAGCTACTTCTCACTTACATTCTGCGTGCAGGTGCAGGGCAAGAGCAAGAGAGAGAAGAAGGATAGAGTGTTCACCGACAAGACCAGCGCCACCGTGATCTGCAGAAAGAACGCCAGCATCTCAGTGAGAGCCCAGGACAGATACTACTCATCCTCCTGGAGCGAGTGGGCCAGCGTGCCTTGCTCCGGTGGTGGTGGCGGAGGCAGCAGAAACCTGCCTGTGGCTACACCTGATCCTGGCATGTTCCCTTGCCTGCACCACAGCCAGAACCTGCTGAGAGCCGTGAGCAACATGCTGCAGAAGGCCAGACAGACTCTGGAGTTCTACCCTTGCACCAGCGAGGAGATCGACCACGAGGACATCACCAAGGATAAGACAAGCACCGTGGAGGCCTGCCTGCCTCTGGAGCTGACCAAGAACGAGAGCTGCCTAAACTCTAGGGAAACCAGCTTCATTACTAACGGCAGTTGCTTAGCCAGCCGGAAGACATCGTTCATGATGGCCCTGTGCCTGAGCAGCATCTACGAGGACCTGAAGATGTACCAAGTGGAGTTCAAGACCATGAACGCCAAGCTGCTGATGGACCCTAAGAGACAGATCTTCCTAGACCAGAACATGCTCGCCGTGATCGACGAGCTGATGCAGGCCCTGAACTTCAACAGCGAAACTGTGCCTCAGAAGAGTTCACTGGAGGAGCCTGACTTCTATAAGACTAAGATCAAGCTGTGTATTCTCCTCCACGCCTTCAGAATCAGGGCTGTCACCATCGATAGGGTGATGAGCTACCTGAACGCATCGTCCGGCGGAGGATCCGGAGGAGGAGGCTCCGGCGGTGGTGGAAGTGGAGGAGGTGGATCAGGAGGCGGTAGTCTCCAGCTCCTGCCTAGCTGGGCCATCACCCTGATCTCTGTAAACGGCATTTTCGTCATTTGCTGTCTGACTTACTGCTTCGCCCCTAGGTGCCGGGAGCGTAGGAGAAACGAGAGACTGCGCCGGGAGTCCGTGCGGCCTGTG 246 h12AB-80TIDATGTGTCACCAGCAGCTCGTGATCAGCTGGTTCAGCCTGGTGTTCCTGGCCTCCCC NucleotideGCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGCTGGACTGGT Sequence 3ACCCCGACGCTCCCGGCGAGATGGTGGTGCTGACCTGCGACACACCGGAGGAAGAC SE_IL12_042GGAATCACCTGGACCCTGGACCAATCCTCCGAAGTTCTGGGCAGCGGCAAGACCCTGACCATCCAGGTGAAGGAGTTCGGCGACGCCGGCCAGTACACCTGCCACAAGGGCGGCGAGGTGCTGAGCCACAGCCTGCTCCTCCTCCACAAGAAAGAGGACGGCATCTGGAGCACCGACATCCTGAAGGACCAGAAGGAGCCCAAGAACAAGACCTTCCTGCGGTGCGAGGCCAAGAACTACTCAGGCCGATTCACCTGTTGGTGGCTCACAACTATCAGCACAGACCTGACCTTCAGCGTGAAGTCTAGCCGGGGCAGCAGCGATCCTCAGGGCGTGACCTGCGGCGCCGCCACCCTGAGCGCCGAGCGGGTGCGGGGCGACAACAAGGAGTACGAGTACAGCGTGGAGTGCCAGGAGGACAGCGCCTGCCCGGCCGCCGAAGAGTCCCTGCCCATCGAGGTGATGGTGGACGCCGTGCACAAACTCAAGTACGAGAACTACACCTCCAGCTTCTTCATCCGGGACATCATCAAGCCCGATCCGCCCAAGAACCTGCAGCTGAAGCCCCTGAAGAACAGCCGGCAGGTGGAGGTGAGCTGGGAATACCCGGACACGTGGTCCACCCCACACAGCTACTTCAGCCTGACCTTTTGCGTGCAGGTCCAAGGCAAGAGCAAGCGGGAGAAGAAGGACCGGGTGTTCACCGATAAGACCTCAGCCACCGTGATTTGCAGAAAGAACGCATCCATATCCGTACGCGCCCAGGATCGGTACTACAGCAGCAGCTGGAGCGAGTGGGCCAGCGTGCCCTGTAGCGGCGGCGGCGGTGGTGGGAGTCGCAACCTGCCCGTGGCCACCCCGGACCCCGGCATGTTCCCCTGCCTGCACCACAGCCAGAACCTGCTGCGGGCCGTGAGCAACATGCTGCAGAAGGCCCGGCAGACCCTGGAGTTCTACCCCTGCACCAGCGAGGAGATCGACCACGAGGACATCACCAAGGACAAGACCAGCACCGTGGAGGCCTGCCTGCCCCTGGAGCTGACCAAGAACGAGAGCTGCCTGAACAGTCGCGAAACCTCCTTCATTACGAACGGCAGCTGCCTGGCCAGCCGGAAGACCAGCTTCATGATGGCCCTGTGCCTGAGCAGCATCTACGAGGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAACGCCAAGCTGCTGATGGACCCCAAGCGGCAGATCTTCCTGGACCAGAACATGCTGGCCGTGATCGACGAGCTGATGCAGGCCCTGAACTTCAACAGCGAAACCGTGCCCCAGAAGTCCAGCCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATCCTGCTGCACGCCTTCCGGATCCGGGCCGTGACCATCGACCGGGTGATGAGCTACCTGAACGCCTCTTCCGGTGGCGGGAGCGGAGGCGGTGGATCTGGCGGAGGAGGGTCGGGAGGCGGCGGAAGCGGTGGTGGAAGCCTTCAACTGCTGCCCTCGTGGGCCATCACACTGATCTCCGTGAACGGCATCTTCGTGATCTGCTGCCTGACCTACTGCTTCGCCCCTCGGTGCCGCGAGCGACGGAGAAACGAGAGGCTCAGACGGGAGAGCGTGCGGCCCGTG 247 h12AB-80TIDATGTGCCACCAACAGCTGGTGATCAGCTGGTTCAGCCTGGTGTTCCTGGCCTCACC NucleotideCCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGCTGGACTGGT Sequence 4ACCCCGACGCCCCTGGCGAGATGGTGGTGCTGACCTGCGACACGCCCGAGGAAGAC SE_IL12_043GGTATCACCTGGACTCTGGACCAGAGCAGCGAGGTGCTGGGCAGCGGCAAGACCCTGACCATCCAGGTGAAGGAGTTCGGCGACGCCGGCCAGTACACCTGCCACAAGGGCGGCGAAGTGCTGAGCCACAGCCTTCTGCTGCTGCACAAGAAGGAGGACGGCATTTGGAGCACCGACATCCTGAAGGACCAGAAGGAGCCCAAGAACAAGACCTTCCTGCGGTGCGAGGCCAAGAACTACAGCGGCCGGTTCACCTGCTGGTGGCTGACCACCATCAGCACCGACCTTACCTTCAGCGTTAAGAGCAGCCGGGGCAGCAGCGATCCCCAGGGCGTGACCTGCGGAGCCGCCACCCTCTCCGCAGAGCGGGTGCGTGGCGACAACAAGGAGTACGAGTACAGCGTGGAGTGCCAGGAGGATAGCGCCTGTCCCGCTGCCGAAGAGAGCCTGCCCATCGAGGTGATGGTGGACGCCGTGCACAAGCTGAAGTACGAGAACTACACCAGCAGCTTCTTCATCCGGGACATCATCAAGCCCGATCCACCCAAGAACCTGCAGCTGAAGCCCCTGAAGAACAGCAGGCAGGTTGAGGTGAGCTGGGAATACCCCGACACCTGGAGCACCCCTCACAGCTACTTCAGCCTGACCTTCTGCGTGCAGGTGCAGGGCAAGAGCAAGCGGGAGAAGAAGGATCGGGTGTTCACCGATAAGACCAGCGCCACCGTGATCTGCCGGAAGAACGCCAGCATCAGCGTTCGGGCCCAGGACCGGTACTACAGCAGCAGCTGGAGCGAGTGGGCCAGCGTGCCCTGCTCTGGAGGCGGAGGCGGAGGCTCACGGAACCTGCCAGTGGCCACGCCGGATCCCGGCATGTTCCCCTGCCTGCACCACAGCCAGAACCTGCTGCGGGCCGTGAGCAACATGCTGCAGAAGGCCCGGCAGACCCTGGAGTTCTACCCCTGCACCAGCGAGGAGATCGACCACGAGGACATCACCAAGGACAAGACCAGCACCGTGGAGGCCTGCCTGCCCCTGGAGCTGACCAAGAACGAGAGCTGCCTGAACAGCCGGGAGACAAGCTTCATCACCAACGGCAGCTGCCTGGCCAGCCGGAAGACCAGCTTCATGATGGCCCTGTGCCTGAGCAGCATCTACGAGGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAACGCCAAGCTGCTGATGGACCCCAAGCGGCAGATCTTCCTGGATCAGAACATGCTGGCCGTGATCGACGAGCTGATGCAGGCCCTGAACTTCAACAGCGAGACTGTGCCCCAGAAGTCCAGCCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATCCTGCTGCACGCCTTCCGGATCCGGGCTGTGACCATCGACCGGGTGATGAGCTACCTGAACGCCTCTTCCGGCGGCGGATCGGGAGGTGGAGGTTCTGGAGGAGGTGGAAGCGGTGGTGGCGGAAGCGGCGGTGGCAGCCTGCAATTGCTCCCCAGCTGGGCCATCACCCTGATCAGCGTGAACGGCATCTTCGTGATCTGTTGCCTGACCTACTGCTTCGCCCCACGGTGCCGGGAGAGACGGCGGAACGAGCGGCTGCGGCGAGAGAGCGTGCGGCCCGTG 248 h12AB-80TIDATGTGTCACCAGCAGTTGGTGATATCTTGGTTCTCACTGGTGTTTCTTGCATCACC NucleotideACTCGTGGCGATCTGGGAACTTAAGAAGGACGTCTACGTGGTGGAGTTAGATTGGT Sequence 5ATCCTGACGCACCCGGGGAAATGGTTGTCCTCACGTGCGACACTCCAGAGGAAGAC SE_IL12_044GGGATCACCTGGACCCTGGATCAGTCGTCAGAGGTACTTGGCAGTGGCAAGACACTGACAATCCAGGTTAAAGAGTTTGGTGACGCCGGGCAGTATACGTGCCACAAGGGCGGCGAGGTGTTGTCACATTCTCTGCTTCTCCTGCACAAGAAAGAAGACGGCATCTGGTCAACTGAGATCCTGAAAGACCAGAAAGAACCCAAGAATAAGACCTTCCTCCGTTGCGAAGCAAAGAACTACTCAGGGCGTTTCACTTGCTGGTGGCTAACCACAATTTCTACCGATCTGACGTTCTCTGTGAAGTCAAGTAGGGGATCCTCAGACCCTCAAGGGGTCACCTGCGGCGCCGCCACCTTATCCGCCGAAAGAGTTCGGGGTGACAATAAAGAGTACGAGTACAGCGTCGAGTGTCAGGAGGACTCCGCCTGTCCTGCTGCAGAGGAGTCCCTGCCGATCGAAGTTATGGTGGACGCCGTCCACAAGCTCAAATACGAGAACTACACCTCAAGCTTCTTCATCAGAGACATCATCAAGCCTGATCCACCCAAGAACCTGCAACTGAAGCCTTTGAAGAACAGCCGACAGGTGGAGGTTTCTTGGGAATATCCAGACACGTGGAGTACGCCCCATTCCTACTTCAGCTTGACCTTCTGCGTGCAGGTTCAGGGGAAGTCCAAGAGAGAGAAGAAGGATCGTGTGTTCACAGACAAGACCTCCGCCACCGTGATCTGCCGGAAGAACGCATCTATCAGTGTTAGGGCCCAGGATCGGTACTACTCGAGTTCCTGGTCTGAGTGGGCAAGTGTGCCCTGCTCCGGTGGCGGCGGAGGAGGGTCAAGGAACCTGCCCGTTGCCACACCAGATCCAGGAATGTTCCCCTGTCTGCACCACTCTCAGAACCTTTTGCGAGCCGTTTCTAATATGCTTCAGAAGGCTCGGCAGACCCTTGAGTTTTATCCCTGCACGTCTGAGGAGATCGATCACGAGGACATCACCAAGGACAAGACTTCCACCGTTGAAGCCTGTTTACCTCTGGAACTGACCAAGAACGAATCCTGTCTCAACAGTAGGGAAACGAGCTTCATCACTAACGGAAGCTGTCTGGCTAGCCGGAAGACCTCTTTTATGATGGCCCTGTGCTTGAGCTCTATTTACGAAGATTTGAAGATGTACCAAGTGGAATTTAAGACTATGAACGCCAAACTGCTGATGGACCCTAAGCGCCAAATCTTCTTGGATCAGAATATGCTGGCTGTAATCGACGAGCTCATGCAGGCTCTGAACTTCAACAGCGAGACGGTACCGCAGAAGAGTTCCCTGGAAGAACCGGACTTCTACAAGACTAAGATTAAACTCTGCATACTCCTCCACGCCTTCCGGATCAGGGCCGTCACAATAGATAGGGTCATGAGTTATCTTAACGCGAGTTCTGGTGGTGGATCGGGTGGCGGAGGCTCAGGAGGAGGCGGTTCTGGCGGTGGTGGGAGTGGAGGCGGTAGTCTGCAGCTGCTGCCGAGTTGGGCAATCACGCTAATCAGCGTGAACGGAATATTCGTAATTTGTTGCCTCACCTATTGTTTCGCACCCAGGTGCAGGGAAAGGAGGCGAAACGAAAGGTTGAGGAGGGAATCTGTCCGGCCAGTG 249 h12AB-80TID MCHQQLVISWFSLVFLASPLVAIWELKKDVYVVELDWYPDAPGEMVVLTCDTPEED Amino AcidGITWTLDQSSEVLGSGKTLTIQVKEFGDAGQYTCHKGGEVLSHSLLLLHKKEDGIW Sequence 1STDILKDQKEPKNKTFLRCEAKNYSGRFTCWWLTTISTDLTFSVKSSRGSSDPQGV (CorrespondsTCGAATLSAERVRGDNKEYEYSVECQEDSACPAAEESLPIEVMVDAVHKLKYENYT to nucleotideSSFFIRDIIKPDPPKNLQLKPLKNSRQVEVSWEYPDTWSTPHSYFSLTFCVQVQGK sequences 1-5)

NLPVATPDPGMFPCLHHSQNLLRAVSNMLQKARQTLEFYPCTSEEIDHEDITKDKTSTVEACLPLELTKNESCLNSRETSFITNGSCLASRKTSFMMALCLSSIYEDLKMYQVEFKTMNAKLLMDPKRQIFLDQNMLAVIDELMQALNFNSETVPQKSSLEEPDFYKT

Italic: signal peptide; Underline: IL12B; Dashed underlineand Italic: linker; Bold: IL12A; Double underline:Transmembrane domain; Bold and Italic: Intracellular domain 250 HumanGUGGUGGUGAUCAGCGCCAUCCUGGCCCUGGUGGUGCUGACCAUCAUCAGCCUGAU PGFRBCAUCCUGAUCAUGCUGUGG transmembrane domain nucleotide sequence 251 HumanCAGAAGAAGCCCAGAUACGAGAUCAGAUGGAAGGUGAUCGAGAGCGUGAGCAGCGA PGFRBCGGCCACGAG E570tr intracellular domain nucleotide sequence 252h12AB-PTM- ATGTGCCACCAGCAGCTGGTGATCAGCTGGTTCAGCCTGGTGTTCCTGGCCAGCCCICD-E570tr CCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGTTGGATTGGTNucleotide ACCCCGACGCCCCCGGCGAGATGGTGGTGCTGACCTGCGACACCCCCGAGGAGGACSequence GGCATCACCTGGACCCTGGACCAGAGCAGCGAGGTGCTGGGCAGCGGCAAGACCCTGACCATCCAGGTGAAGGAGTTCGGCGACGCCGGCCAGTACACCTGCCACAAGGGCGGCGAGGTGCTGAGCCACAGCCTGCTGCTGCTGCACAAGAAGGAGGACGGCATCTGGAGCACCGACATCCTGAAGGACCAGAAGGAGCCCAAGAACAAGACCTTCCTGAGATGCGAGGCCAAGAACTACAGCGGCAGATTCACCTGCTGGTGGCTGACCACCATCAGCACCGACCTGACCTTCAGCGTGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGTGACCTGCGGCGCCGCCACCCTGAGCGCCGAGAGAGTGAGAGGCGACAACAAGGAGTACGAGTACAGCGTGGAGTGCCAGGAAGATAGCGCCTGCCCCGCCGCCGAGGAGAGCCTGCCCATCGAGGTGATGGTGGACGCCGTGCACAAGCTGAAGTACGAGAACTACACCAGCAGCTTCTTCATCAGAGATATCATCAAGCCCGACCCCCCCAAGAACCTGCAGCTGAAGCCCCTGAAGAACAGCCGGCAGGTGGAGGTGAGCTGGGAGTACCCCGACACCTGGAGCACCCCCCACAGCTACTTCAGCCTGACCTTCTGCGTGCAGGTGCAGGGCAAGAGCAAGAGAGAGAAGAAAGATAGAGTGTTCACCGACAAGACCAGCGCCACCGTGATCTGCAGAAAGAACGCCAGCATCAGCGTGAGAGCCCAAGATAGATACTACAGCAGCAGCTGGAGCGAGTGGGCCAGCGTGCCCTGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCTGCCCGTGGCCACCCCCGACCCCGGCATGTTCCCCTGCCTGCACCACAGCCAGAACCTGCTGAGAGCCGTGAGCAACATGCTGCAGAAGGCCCGGCAGACCCTGGAGTTCTACCCCTGCACCAGCGAGGAGATCGACCACGAAGATATCACCAAAGATAAGACCAGCACCGTGGAGGCCTGCCTGCCCCTGGAGCTGACCAAGAACGAGAGCTGCCTGAACAGCAGAGAGACCAGCTTCATCACCAACGGCAGCTGCCTGGCCAGCAGAAAGACCAGCTTCATGATGGCCCTGTGCCTGAGCAGCATCTACGAGGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAACGCCAAGCTGCTGATGGACCCCAAGCGGCAGATCTTCCTGGACCAGAACATGCTGGCCGTGATCGACGAGCTGATGCAGGCCCTGAACTTCAACAGCGAGACCGTGCCCCAGAAGAGCAGCCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATCCTGCTGCACGCCTTCAGAATCAGAGCCGTGACCATCGACAGAGTGATGAGCTACCTGAACGCCAGCTCTGGTGGCGGATCAGGCGGCGGCGGTTCAGGAGGCGGTGGAAGTGGAGGTGGCGGGTCTGGCGGAGGTTCACTGCAGGTGGTGGTGATCAGCGCCATCCTGGCCCTGGTGGTGCTCACCATCATCAGCCTGATCATCCTGATCATGCTGTGGCAGAAGAAGCCCAGATACGAGATCCGGTGGAAGGTGATCGAGAGCGTGAGCAGCGACGGCCACGAGTTCATCTTCGTGGACCCCATGCAGCTGCCCTACGACAGCACCTGGGAGCTGCCCCGTGATCAGCTGGTGCTGGGCAGAACCCTGGGCAGCGGCGCCTTCGGCCAGGTGGTGGAGGCTACCGCCCACGGCCTGAGCCACAGCCAGGCCACCATGAAGGTGGCCGTGGCCATGCTCAAGAGCACCGCCAGAAGCAGCGAGAAGCAGGCCCTGATGAGCGAGCTGAAGATCATGAGCCATCTGGGGCCCCACCTGAACGTGGTGAACCTGCTGGGCGCCTGCACCAAGGGCGGCCCCATCTACATCATCACCGAGTACTGCAGATACGGCGACCTGGTGGACTACCTGCACAGAAACAAGCACACCTTCCTGCAGCACCACAGCGACAAGAGAAGACCTCCCAGCGCCGAGCTGTACAGCAACGCCCTGCCCGTTGGTCTGCCCCTACCCAGCCACGTGAGCCTGACCGGCGAGAGCGACGGCG GC 253h12AB-PTM- MCHQQLVISWFSLVFLASPLVA IWELKKDVYVVELDWYPDAPGEMVVLTCDTPEEDICD-E570tr GITWTLDQSSEVLGSGKTLTIQVKEFGDAGQYTCHKGGEVLSHSLLLLHKKEDGIWAmino Acid STDILKDQKEPKNKTFLRCEAKNYSGRFTCWWLTTISTDLTFSVKSSRGSSDPQGVSequence TCGAATLSAERVRGDNKEYEYSVECQEDSACPAAEESLPIEVMVDAVHKLKYENYTSSFFIRDIIKPDPPKNLQLKPLKNSRQVEVSWEYPDTWSTPHSYFSLTFCVQVQGK

RNLPVATPDPGMFPCLHHSQNLLRAVSNMLQKARQTLEFYPCTSEEIDHEDITKDKTSTVEACLPLELTKNESCLNSRETSFITNGSCLASRKTSFMMALCLSSIYEDLKMYQVEFKTMNAKLLMDPKRQIFLDQNMLAVIDELMQALNFNSETVPQKSSLEEPDFYKT

Italic: signal peptide; Underline: IL12B; Dashed underlineand Italic: linker; Bold: IL12A; Double underline:Transmembrane domain; Bold and Italic: Intracellular domain 254h12AB-PTM- ATGTGCCACCAGCAGCTGGTGATCAGCTGGTTCAGCCTGGTGTTCCTGGCCAGCCCICD-G739tr CCTGGTGGCCATCTGGGAGCTGAAGAAGGACGTGTACGTGGTGGAGTTGGATTGGTNucleotide ACCCCGACGCCCCCGGCGAGATGGTGGTGCTGACCTGCGACACCCCCGAGGAGGACSequence GGCATCACCTGGACCCTGGACCAGAGCAGCGAGGTGCTGGGCAGCGGCAAGACCCTGACCATCCAGGTGAAGGAGTTCGGCGACGCCGGCCAGTACACCTGCCACAAGGGCGGCGAGGTGCTGAGCCACAGCCTGCTGCTGCTGCACAAGAAGGAGGACGGCATCTGGAGCACCGACATCCTGAAGGACCAGAAGGAGCCCAAGAACAAGACCTTCCTGAGATGCGAGGCCAAGAACTACAGCGGCAGATTCACCTGCTGGTGGCTGACCACCATCAGCACCGACCTGACCTTCAGCGTGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGTGACCTGCGGCGCCGCCACCCTGAGCGCCGAGAGAGTGAGAGGCGACAACAAGGAGTACGAGTACAGCGTGGAGTGCCAGGAAGATAGCGCCTGCCCCGCCGCCGAGGAGAGCCTGCCCATCGAGGTGATGGTGGACGCCGTGCACAAGCTGAAGTACGAGAACTACACCAGCAGCTTCTTCATCAGAGATATCATCAAGCCCGACCCCCCCAAGAACCTGCAGCTGAAGCCCCTGAAGAACAGCCGGCAGGTGGAGGTGAGCTGGGAGTACCCCGACACCTGGAGCACCCCCCACAGCTACTTCAGCCTGACCTTCTGCGTGCAGGTGCAGGGCAAGAGCAAGAGAGAGAAGAAAGATAGAGTGTTCACCGACAAGACCAGCGCCACCGTGATCTGCAGAAAGAACGCCAGCATCAGCGTGAGAGCCCAAGATAGATACTACAGCAGCAGCTGGAGCGAGTGGGCCAGCGTGCCCTGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCTGCCCGTGGCCACCCCCGACCCCGGCATGTTCCCCTGCCTGCACCACAGCCAGAACCTGCTGAGAGCCGTGAGCAACATGCTGCAGAAGGCCCGGCAGACCCTGGAGTTCTACCCCTGCACCAGCGAGGAGATCGACCACGAAGATATCACCAAAGATAAGACCAGCACCGTGGAGGCCTGCCTGCCCCTGGAGCTGACCAAGAACGAGAGCTGCCTGAACAGCAGAGAGACCAGCTTCATCACCAACGGCAGCTGCCTGGCCAGCAGAAAGACCAGCTTCATGATGGCCCTGTGCCTGAGCAGCATCTACGAGGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAACGCCAAGCTGCTGATGGACCCCAAGCGGCAGATCTTCCTGGACCAGAACATGCTGGCCGTGATCGACGAGCTGATGCAGGCCCTGAACTTCAACAGCGAGACCGTGCCCCAGAAGAGCAGCCTGGAGGAGCCCGACTTCTACAAGACCAAGATCAAGCTGTGCATCCTGCTGCACGCCTTCAGAATCAGAGCCGTGACCATCGACAGAGTGATGAGCTACCTGAACGCCAGCTCTGGTGGCGGATCAGGCGGCGGCGGTTCAGGAGGCGGTGGAAGTGGAGGTGGCGGGTCTGGCGGAGGTTCACTGCAGGTGGTGGTGATCAGCGCCATCCTGGCCCTGGTGGTGCTCACCATCATCAGCCTGATCATCCTGATCATGCTGTGGCAGAAGAAGCCCAGATACGAGATCCGGTGGAAGGTGATCGAGAGCGTGAGCAGCGACGGCCACGAGTTCATCTTCGTGGACCCCATGCAGCTGCCCTACGACAGCACCTGGGAGCTGCCCCGTGATCAGCTGGTGCTGGGCAGAACCCTGGGCAGCGGCGCCTTCGGCCAGGTGGTGGAGGCTACCGCCCACGGCCTGAGCCACAGCCAGGCCACCATGAAGGTGGCCGTGGCCATGCTCAAGAGCACCGCCAGAAGCAGCGAGAAGCAGGCCCTGATGAGCGAGCTGAAGATCATGAGCCATCTGGGGCCCCACCTGAACGTGGTGAACCTGCTGGGCGCCTGCACCAAGGGCGGCCCCATCTACATCATCACCGAGTACTGCAGATACGGCGACCTGGTGGACTACCTGCACAGAAACAAGCACACCTTCCTGCAGCACCACAGCGACAAGAGAAGACCTCCCAGCGCCGAGCTGTACAGCAACGCCCTGCCCGTTGGTCTGCCCCTACCCAGCCACGTGAGCCTGACCGGCGAGAGCGACGGCG GC 255h12AB-PTM- MCHQQLVISWFSLVFLASPLVA IWELKKDVYVVELDWYPDAPGEMVVLTCDTPEEDICD-G739tr GITWTLDQSSEVLGSGKTLTIQVKEFGDAGQYTCHKGGEVLSHSLLLLHKKEDGIWAmino Acid STDILKDQKEPKNKTFLRCEAKNYSGRFTCWWLTTISTDLTFSVKSSRGSSDPQGVSequence TCGAATLSAERVRGDNKEYEYSVECQEDSACPAAEESLPIEVMVDAVHKLKYENYTSSFFIRDIIKPDPPKNLQLKPLKNSRQVEVSWEYPDTWSTPHSYFSLTFCVQVQGK

RNLPVATPDPGMFPCLHHSQNLLRAVSNMLQKARQTLEFYPCTSEEIDHEDITKDKTSTVEACLPLELTKNESCLNSRETSFITNGSCLASRKTSFMMALCLSSIYEDLKMYQVEFKTMNAKLLMDPKRQIFLDQNMLAVIDELMQALNFNSETVPQKSSLEEPDFYKT

 

Italic: signal peptide; Underline: IL12B; Dashed underlineand Italic: linker; Bold: IL12A; Double underline:Transmembrane domain; Bold and Italic: Intracellular domain 256 [CCG]₄CCGCCGCCGCCG 257 [CCG]₅ CCGCCGCCGCCGCCG 258 V1 GC-rich CCCCGGCGCCRNA element 259 V2 GC-rich CCCCGGC RNA element 260 EK GC-rich GCCGCCRNA element 261 5′UTR GGGAAATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGA 262V1-5′UTR GGGAAATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGACCCCGGCGCCGCCACC 263V2-5′UTR GGGAAATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGACCCCGGCGCCACC 264Standard GGGAAATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACC 5′UTR 265 KozakGCCA/GCC consensus 266 3′UTR withTGATAATAGGCTGGAGCCTCGGTGGCCTAGCTTCTTGCCCCTTGGGCCTCCCCCCA mir-122-5pGCCCCTCCTCCCCTTCCTGCACCCGTACCCCC 

GT binding site GGTCTTTGAATAAAGTCTGAGTGGGCGGC 267 mIL12AB-04-GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGUCCU 001CAGAAGCUAACCAUCUCCUGGUUUGCCAUCGUUUUGCUGGUGUCUCCACUCAUGGCCAUGUGGGAGCUGGAGAAAGACGUUUAUGUUGUAGAGGUGGACUGGACUCCCGAUGCCCCUGGAGAAACAGUGAACCUCACCUGUGACACGCCUGAAGAAGAUGACAUCACCUGGACCUCAGACCAGAGACAUGGAGUCAUAGGCUCUGGAAAGACCCUGACCAUCACUGUCAAAGAGUUCCUAGAUGCUGGCCAGUACACCUGCCACAAAGGAGGCGAGACUCUGAGCCACUCACAUCUGCUGCUCCACAAGAAGGAAAAUGGAAUUUGGUCCACUGAAAUUUUAAAAAAUUUCAAAAACAAGACUUUCCUGAAGUGUGAAGCACCAAAUUACUCCGGACGGUUCACGUGCUCAUGGCUGGUGCAAAGAAACAUGGACUUGAAGUUCAACAUCAAGAGCAGUAGCAGUUCCCCUGACUCUCGGGCAGUGACAUGUGGAAUGGCGUCUCUGUCUGCAGAGAAGGUCACACUGGACCAAAGGGACUAUGAGAAGUAUUCAGUGUCCUGCCAGGAGGAUGUCACCUGCCCAACUGCCGAGGAGACCCUGCCCAUUGAACUGGCGUUGGAAGCACGGCAGCAGAAUAAAUAUGAGAACUACAGCACCAGCUUCUUCAUCAGGGACAUCAUCAAACCAGACCCGCCCAAGAACUUGCAGAUGAAGCCUUUGAAGAACUCACAGGUGGAGGUCAGCUGGGAGUACCCUGACUCCUGGAGCACUCCCCAUUCCUACUUCUCCCUCAAGUUCUUUGUUCGAAUCCAGCGCAAGAAAGAAAAGAUGAAGGAGACAGAGGAGGGGUGUAACCAGAAAGGUGCGUUCCUCGUAGAGAAGACAUCUACCGAAGUCCAAUGCAAAGGCGGGAAUGUCUGCGUGCAAGCUCAGGAUCGCUAUUACAAUUCCUCAUGCAGCAAGUGGGCAUGUGUUCCCUGCAGGGUCCGAUCCGGAGGCGGAGGGAGCGGAGGCGGAGGGAGCGGAGGCGGAGGGAGCAGGGUCAUUCCAGUCUCUGGACCUGCCAGGUGUCUUAGCCAGUCCCGAAACCUGCUGAAGACCACAGAUGACAUGGUGAAGACGGCCAGAGAAAAACUGAAACAUUAUUCCUGCACUGCUGAAGACAUCGAUCAUGAAGACAUCACACGGGACCAAACCAGCACAUUGAAGACCUGUUUACCACUGGAACUACACAAGAACGAGAGUUGCCUGGCUACUAGAGAGACUUCUUCCACAACAAGAGGGAGCUGCCUGCCCCCACAGAAGACGUCUUUGAUGAUGACCCUGUGCCUUGGUAGCAUCUAUGAGGACUUGAAGAUGUACCAGACAGAGUUCCAGGCCAUCAACGCAGCACUUCAGAAUCACAACCAUCAGCAGAUCAUUUUAGACAAGGGCAUGCUGGUGGCCAUCGAUGAGCUGAUGCAGUCUCUGAAUCAUAAUGGCGAGACUCUGCGCCAGAAACCUCCUGUGGGAGAAGCAGACCCUUACAGAGUGAAAAUGAAGCUCUGCAUCCUGCUUCACGCCUUCAGCACCCGCGUCGUGACCAUCAACAGGGUGAUGGGCUAUCUGAGCUCCGCCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 268 mIL12ABAUGUGUCCUCAGAAGCUAACCAUCUCCUGGUUUGCCAUCGUUUUGUUGGUGUCUCC 80TM-ACUCAUGGCCAUGUGGGAGCUCGAGAAAGACGUUUACGUUGUAGAGGUGGACUGGA nolinker_CUCCCGACGCCCCGGGCGAAACAGUGAACCUCACCUGUGACACGCCUGAAGAAGAU V5_GACAUCACCUGGACCUCAGACCAGAGACAUGGAGUCAUAGGCUCUGGAAAGACCCU NucleotideGACCAUCACUGUCAAAGAGUUCCUAGAUGCUGGCCAGUACACCUGCCACAAAGGAG SequenceGCGAGACUCUGAGCCACUCACAUCUGCUGCUCCACAAGAAGGAGAAUGGAAUUUGGUCCACUGAGAUCCUGAAGAACUUCAAGAAUAAGACUUUCCUGAAGUGUGAAGCACCAAAUUACUCCGGACGGUUCACGUGCUCAUGGCUGGUGCAAAGAAACAUGGACUUGAAGUUCAACAUCAAGAGCAGUAGCAGUUCCCCUGACUCUCGGGCAGUGACAUGUGGAAUGGCGUCUCUGUCUGCAGAGAAGGUCACACUGGACCAAAGGGACUAUGAGAAGUAUUCAGUGUCCUGCCAGGAGGAUGUCACCUGCCCAACUGCCGAGGAGACACUGCCCAUUGAACUGGCGUUGGAAGCACGGCAGCAGAAUAAAUAUGAGAACUACAGCACCAGCUUCUUCAUCAGGGACAUCAUCAAACCAGACCCGCCCAAGAACUUGCAGAUGAAGCCUUUGAAGAACUCACAGGUGGAGGUCAGCUGGGAGUACCCGGACUCCUGGAGCACUCCCCAUUCCUACUUCUCCCUCAAGUUCUUUGUUCGAAUCCAGCGCAAGAAAGAGAAGAUGAAGGAGACAGAGGAGGGGUGUAACCAGAAAGGUGCGUUCCUCGUAGAGAAGACAUCUACCGAAGUCCAAUGCAAAGGCGGGAAUGUCUGCGUGCAAGCUCAGGAUCGCUAUUACAAUUCCUCAUGCAGCAAGUGGGCAUGUGUUCCCUGCAGGGUCCGAUCCGGAGGCGGAGGGAGCGGUGGUGGAGGCAGCGGAGGAGGUGGAUCAAGGGUCAUUCCAGUCUCUGGACCAGCUAGAUGUCUUAGCCAGUCCCGAAACCUGCUGAAGACCACGGACGACAUGGUGAAGACGGCCAGAGAGAAACUGAAACAUUAUUCCUGCACCGCAGAGGAUAUCGAUCACGAAGAUAUCACACGGGACCAAACCAGCACAUUGAAGACCUGUUUACCACUGGAACUACACAAGAACGAGAGUUGCCUGGCUACUAGAGAGACUUCUUCCACAACAAGAGGGAGCUGCCUGCCACCACAGAAGACGUCUUUGAUGAUGACCCUGUGCCUUGGUAGCAUCUAUGAGGACCUCAAGAUGUACCAGACAGAGUUCCAGGCCAUCAACGCAGCACUUCAGAAUCACAACCAUCAGCAGAUCAUUUUAGACAAGGGCAUGCUGGUGGCCAUCGAUGAGCUGAUGCAAUCAUUGAAUCAUAACGGUGAGACAUUGCGCCAGAAACCUCCUGUGGGAGAAGCAGACCCUUACAGAGUGAAGAUGAAGCUCUGCAUCCUGCUUCACGCCUUCAGCACCCGCGUCGUCACUAUCAACAGGGUGAUGGGCUAUCUGAGCUCCGCCACCCUGGUGCUGUUCGGCGCCGGCUUCGGUGCAGUGAUCACCGUGGUGGUGAUCGUCGUCAUCAUCGGGAAACCAAUUCCAAAUCCCCUCCUGGGGUUGGAUAGCACC 269 mIL12AB-AUGUGUCCUCAGAAGCUAACCAUCUCCUGGUUUGCCAUCGUUUUGCUUGUGUCUCC PTM_v5ACUCAUGGCCAUGUGGGAGCUCGAGAAAGACGUUUACGUUGUAGAGGUGGACUGGA miR122CUCCCGACGCCCCAGGAGAAACAGUGAACCUCACCUGUGACACGCCUGAAGAAGAU NucleotideGACAUCACCUGGACCUCAGACCAGAGACAUGGAGUCAUAGGCUCUGGAAAGACCCU SequenceGACCAUCACUGUCAAAGAGUUCCUAGAUGCUGGCCAGUACACCUGCCACAAAGGAGGCGAGACUCUGAGCCACUCACAUCUGCUGCUCCACAAGAAGGAGAAUGGAAUUUGGUCCACCGAAAUCCUGAAGAACUUCAAGAAUAAGACUUUCCUGAAGUGUGAAGCACCAAAUUACUCCGGACGGUUCACGUGCUCAUGGCUGGUGCAAAGAAACAUGGACUUGAAGUUCAACAUCAAGAGCAGUAGCAGUUCCCCUGACUCUCGGGCAGUGACAUGUGGAAUGGCGUCUCUGUCUGCAGAGAAGGUCACACUGGACCAAAGGGACUAUGAGAAGUAUUCAGUGUCCUGCCAGGAGGAUGUCACCUGCCCAACUGCCGAGGAAACUCUGCCCAUUGAACUGGCGUUGGAAGCACGGCAGCAGAAUAAAUAUGAGAACUACAGCACCAGCUUCUUCAUCAGGGACAUCAUCAAACCAGACCCGCCCAAGAACUUGCAGAUGAAGCCUUUGAAGAACUCACAGGUGGAGGUCAGCUGGGAGUACCCAGACUCCUGGAGCACUCCCCAUUCCUACUUCUCCCUCAAGUUCUUUGUUCGAAUCCAGCGCAAGAAAGAGAAGAUGAAGGAGACAGAGGAGGGGUGUAACCAGAAAGGUGCGUUCCUCGUAGAGAAGACAUCUACCGAAGUCCAAUGCAAAGGCGGGAAUGUCUGCGUGCAAGCUCAGGAUCGCUAUUACAAUUCCUCAUGCAGCAAGUGGGCAUGUGUUCCCUGCAGGGUCCGAUCCGGAGGCGGAGGGUCUGGAGGAGGAGGUUCUGGAGGUGGUGGCAGUAGGGUCAUUCCAGUCUCUGGACCUGCAAGGUGUCUUAGCCAGUCCCGAAACCUGCUGAAGACCACAGACGAUAUGGUGAAGACGGCCAGAGAGAAACUGAAACAUUAUUCCUGCACAGCAGAGGACAUCGAUCAUGAAGAUAUUACACGGGACCAAACCAGCACAUUGAAGACCUGUUUACCACUGGAACUACACAAGAACGAGAGUUGCCUGGCUACUAGAGAGACUUCUUCCACAACAAGAGGGAGCUGCCUGCCACCACAGAAGACGUCUUUGAUGAUGACCCUGUGCCUUGGUAGCAUCUACGAGGAUCUCAAGAUGUACCAGACAGAGUUCCAGGCCAUCAACGCAGCACUUCAGAAUCACAACCAUCAGCAGAUCAUUUUAGACAAGGGCAUGCUGGUGGCCAUCGAUGAGCUGAUGCAAUCUCUGAAUCAUAAUGGCGAGACACUUCGCCAGAAACCUCCUGUGGGAGAAGCAGACCCUUACAGAGUGAAGAUGAAGCUCUGCAUCCUGCUUCACGCCUUCAGCACCCGCGUCGUCACUAUUAACAGGGUGAUGGGCUAUCUGAGCUCCGCCUCUGGUGGCGGAUCAGGCGGCGGCGGCUCUGGCGGCGGUGGAAGCGGAGGUGGCGGGUCUGGCGGAGGUUCACUGCAGGUAGUAGUGAUCAGCGCCAUCCUGGCCCUGGUGGUGCUGACCGUGAUCUCAUUGAUCAUCUUGAUUAUGCUGUGGGGCGGAGGAGGCAGCGGGAAACCAAUUCCAAAUCCCCUCCUGGGGUUGGAUAGCACC 270 mIL12AB-AUGUGUCCUCAGAAGCUAACCAUCUCCUGGUUUGCCAUCGUUUUGUUAGUGUCUCC 8TM_v5_miRACUCAUGGCCAUGUGGGAGCUCGAGAAAGACGUUUACGUUGUAGAGGUGGACUGGA 122CUCCCGACGCCCCAGGCGAAACAGUGAACCUCACCUGUGACACGCCUGAAGAAGAU NucleotideGACAUCACCUGGACCUCAGACCAGAGACAUGGAGUCAUAGGCUCUGGAAAGACCCU sequenceGACCAUCACUGUCAAAGAGUUCCUAGAUGCUGGCCAGUACACCUGCCACAAAGGAGGCGAGACUCUGAGCCACUCACAUCUGCUGCUCCACAAGAAGGAGAAUGGAAUUUGGUCCACAGAAAUUUUAAAGAACUUCAAGAACAAGACUUUCCUGAAGUGUGAAGCACCAAAUUACUCCGGACGGUUCACGUGCUCAUGGCUGGUGCAAAGAAACAUGGACUUGAAGUUCAACAUCAAGAGCAGUAGCAGUUCCCCUGACUCUCGGGCAGUGACAUGUGGAAUGGCGUCUCUGUCUGCAGAGAAGGUCACACUGGACCAAAGGGACUAUGAGAAGUAUUCAGUGUCCUGCCAGGAGGAUGUCACCUGCCCAACUGCCGAGGAGACUCUGCCCAUUGAACUGGCGUUGGAAGCACGGCAGCAGAAUAAAUAUGAGAACUACAGCACCAGCUUCUUCAUCAGGGACAUCAUCAAACCAGACCCGCCCAAGAACUUGCAGAUGAAGCCUUUGAAGAACUCACAGGUGGAGGUCAGCUGGGAGUACCCAGACUCCUGGAGCACUCCCCAUUCCUACUUCUCCCUCAAGUUCUUUGUUGGAAUCCAGGGCAAGAAAGAGAAGAUGAAGGAGACAGAGGAGGGGUGUAACCAGAAAGGUGCGUUCCUCGUAGAGAAGACAUCUACCGAAGUCCAAUGCAAAGGCGGGAAUGUCUGCGUGCAAGCUCAGGAUCGCUAUUACAAUUCCUCAUGCAGCAAGUGGGCAUGUGUUCCCUGCAGGGUCCGAUCCGGAGGCGGAGGGAGUGGAGGAGGUGGCUCUGGCGGCGGUGGAAGUAGGGUCAUUCCAGUCUCUGGACCUGCACGCUGUCUUAGCCAGUCCCGAAACCUGCUGAAGACCACAGACGACAUGGUGAAGACGGCCAGAGAGAAACUGAAACAUUAUUCCUGCACAGCGGAAGACAUAGAUCACGAGGAUAUCACACGGGACCAAACCAGCACAUUGAAGACCUGUUUACCACUGGAACUACACAAGAACGAGAGUUGCCUGGCUACUAGAGAGACUUCUUCCACAACAAGAGGGAGCUGCCUGCCACCACAGAAGACGUCUUUGAUGAUGACCCUGUGCCUUGGUAGCAUCUAUGAGGAUCUGAAGAUGUACCAGACAGAGUUCCAGGCCAUCAACGCAGCACUUCAGAAUCACAACCAUCAGCAGAUCAUUUUAGACAAGGGCAUGCUGGUGGCCAUCGAUGAGCUGAUGCAGUCCCUGAAUCAUAAUGGUGAAACGUUGCGCCAGAAACCUCCUGUGGGAGAAGCAGACCCUUACAGAGUGAAGAUGAAGCUCUGCAUCCUGCUUCACGCCUUCAGCACCCGCGUCGUGACUAUAAACAGGGUGAUGGGCUAUCUGAGCUCCGCCUCUGGUGGCGGAUCAGGAGGAGGUGGAUCCGGUGGCGGUGGUUCCGGAGGUGGUGGAUCGGGUGGUGGCUCACUGCAGAUCUACAUCUGGGCCCCGCUGGCCGGCAUCUGCGUGGCCCUGCUGCUGAGCCUGAUCAUCACCCUGAUCUGCUACGGUGGAGGCGGUAGCGGGAAACCAAUUCCAAAUCCCCUCCUGGGGUUGGAUAGCACC 271 mIL12AB-AUGUGUCCUCAGAAGCUAACCAUCUCCUGGUUUGCCAUCGUUUUGUUGGUGUCUCC 80TM-ICDACUCAUGGCCAUGUGGGAGCUCGAGAAAGACGUUUACGUUGUAGAGGUGGACUGGA NucleotideCUCCCGACGCCCCGGGCGAAACAGUGAACCUCACCUGUGACACGCCUGAAGAAGAU SequenceGACAUCACCUGGACCUCAGACCAGAGACAUGGAGUCAUAGGCUCUGGAAAGACCCUGACCAUCACUGUCAAAGAGUUCCUAGAUGCUGGCCAGUACACCUGCCACAAAGGAGGCGAGACUCUGAGCCACUCACAUCUGCUGCUCCACAAGAAGGAGAAUGGAAUUUGGUCCACUGAGAUCCUGAAGAACUUCAAGAAUAAGACUUUCCUGAAGUGUGAAGCACCAAAUUACUCCGGACGGUUCACGUGCUCAUGGCUGGUGCAAAGAAACAUGGACUUGAAGUUCAACAUCAAGAGCAGUAGCAGUUCCCCUGACUCUCGGGCAGUGACAUGUGGAAUGGCGUCUCUGUCUGCAGAGAAGGUCACACUGGACCAAAGGGACUAUGAGAAGUAUUCAGUGUCCUGCCAGGAGGAUGUCACCUGCCCAACUGCCGAGGAGACACUGCCCAUUGAACUGGCGUUGGAAGCACGGCAGCAGAAUAAAUAUGAGAACUACAGCACCAGCUUCUUCAUCAGGGACAUCAUCAAACCAGACCCGCCCAAGAACUUGCAGAUGAAGCCUUUGAAGAACUCACAGGUGGAGGUCAGCUGGGAGUACCCGGACUCCUGGAGCACUCCCCAUUCCUACUUCUCCCUCAAGUUCUUUGUUCGAAUCCAGCGCAAGAAAGAGAAGAUGAAGGAGACAGAGGAGGGGUGUAACCAGAAAGGUGCGUUCCUCGUAGAGAAGACAUCUACCGAAGUCCAAUGCAAAGGCGGGAAUGUCUGCGUGCAAGCUCAGGAUCGCUAUUACAAUUCCUCAUGCAGCAAGUGGGCAUGUGUUCCCUGCAGGGUCCGAUCCGGAGGCGGAGGGAGCGGUGGUGGAGGCAGCGGAGGAGGUGGAUCAAGGGUCAUUCCAGUCUCUGGACCAGCUAGAUGUCUUAGCCAGUCCCGAAACCUGCUGAAGACCACGGACGACAUGGUGAAGACGGCCAGAGAGAAACUGAAACAUUAUUCCUGCACCGCAGAGGAUAUCGAUCACGAAGAUAUCACACGGGACCAAACCAGCACAUUGAAGACCUGUUUACCACUGGAACUACACAAGAACGAGAGUUGCCUGGCUACUAGAGAGACUUCUUCCACAACAAGAGGGAGCUGCCUGCCACCACAGAAGACGUCUUUGAUGAUGACCCUGUGCCUUGGUAGCAUCUAUGAGGACCUCAAGAUGUACCAGACAGAGUUCCAGGCCAUCAACGCAGCACUUCAGAAUCACAACCAUCAGCAGAUCAUUUUAGACAAGGGCAUGCUGGUGGCCAUCGAUGAGCUGAUGCAAUCAUUGAAUCAUAACGGUGAGACAUUGCGCCAGAAACCUCCUGUGGGAGAAGCAGACCCUUACAGAGUGAAGAUGAAGCUCUGCAUCCUGCUUCACGCCUUCAGCACCCGCGUCGUCACUAUCAACAGGGUGAUGGGCUAUCUGAGCUCCGCCAGCGGUGGCGGAAGCGGUGGAGGCGGCAGCGGCGGUGGUGGUAGCGGCGGCGGCGGCUCCGGCGGAGGGAGCCUGCAGACCCUGGUGCUGUUCGGCGCCGGCUUCGGUGCAGUGAUCACCGUGGUGGUGAUCGUCGUCAUCAUCAAGUGCUUCUGCAAGCACAGAAGCUGCUUCAGAAGAAACGAGGCCAGCAGAGAAACCAACAACAGCCUAACAUUCGGCCCAGAAGAGGCUCUGGCC 272 IgK-AUGGAGACUGACACCCUGCUGCUGUGGGUGCUGUUACUUUGGGUUCCCGGCAGCAC mscIL12AB-CGGCUACCCCUACGACGUGCCCGACUACGCCAUGUGGGAGCUCGAGAAAGACGUUU 80TM-ICDACGUUGUAGAGGUGGACUGGACUCCCGACGCCCCGGGCGAAACAGUGAACCUCACC NucleotideUGUGACACGCCUGAAGAAGAUGACAUCACCUGGACCUCAGACCAGAGACAUGGAGU SequenceCAUAGGCUCUGGAAAGACCCUGACCAUCACUGUCAAAGAGUUCCUAGAUGCUGGCCAGUACACCUGCCACAAAGGAGGCGAGACUCUGAGCCACUCACAUCUGCUGCUCCACAAGAAGGAGAAUGGAAUUUGGUCCACUGAGAUCCUGAAGAACUUCAAGAAUAAGACUUUCCUGAAGUGUGAAGCACCAAAUUACUCCGGACGGUUCACGUGCUCAUGGCUGGUGCAAAGAAACAUGGACUUGAAGUUCAACAUCAAGAGCAGUAGCAGUUCCCCUGACUCUCGGGCAGUGACAUGUGGAAUGGCGUCUCUGUCUGCAGAGAAGGUCACACUGGACCAAAGGGACUAUGAGAAGUAUUCAGUGUCCUGCCAGGAGGAUGUCACCUGCCCAACUGCCGAGGAGACACUGCCCAUUGAACUGGCGUUGGAAGCACGGCAGCAGAAUAAAUAUGAGAACUACAGCACCAGCUUCUUCAUCAGGGACAUCAUCAAACCAGACCCGCCCAAGAACUUGCAGAUGAAGCCUUUGAAGAACUCACAGGUGGAGGUCAGCUGGGAGUACCCGGACUCCUGGAGCACUCCCCAUUCCUACUUCUCCCUCAAGUUCUUUGUUCGAAUCCAGCGCAAGAAAGAGAAGAUGAAGGAGACAGAGGAGGGGUGUAACCAGAAAGGUGCGUUCCUCGUAGAGAAGACAUCUACCGAAGUCCAAUGCAAAGGCGGGAAUGUCUGCGUGCAAGCUCAGGAUCGCUAUUACAAUUCCUCAUGCAGCAAGUGGGCAUGUGUUCCCUGCAGGGUCCGAGGAAGCACCAGCGGUUCCGGCAAACCAGGUAGCGGAGAGGGCAGCACCAAGGGCAGGGUCAUUCCAGUCUCUGGACCAGCUAGAUGUCUUAGCCAGUCCCGAAACCUGCUGAAGACCACGGACGACAUGGUGAAGACGGCCAGAGAGAAACUGAAACAUUAUUCCUGCACCGCAGAGGAUAUCGAUCACGAAGAUAUCACACGGGACCAAACCAGCACAUUGAAGACCUGUUUACCACUGGAACUACACAAGAACGAGAGUUGCCUGGCUACUAGAGAGACUUCUUCCACAACAAGAGGGAGCUGCCUGCCACCACAGAAGACGUCUUUGAUGAUGACCCUGUGCCUUGGUAGCAUCUAUGAGGACCUCAAGAUGUACCAGACAGAGUUCCAGGCCAUCAACGCAGCACUUCAGAAUCACAACCAUCAGCAGAUCAUUUUAGACAAGGGCAUGCUGGUGGCCAUCGAUGAGCUGAUGCAAUCAUUGAAUCAUAACGGUGAGACAUUGCGCCAGAAACCUCCUGUGGGAGAAGCAGACCCUUACAGAGUGAAGAUGAAGCUCUGCAUCCUGCUUCACGCCUUCAGCACCCGCGUCGUCACUAUCAACAGGGUGAUGGGCUAUCUGAGCUCCGCCACCCUGGUGCUGUUCGGCGCCGGCUUCGGUGCAGUGAUCACCGUGGUGGUGAUCGUCGUCAUCAUCAAGUGCUUUUGCAAGCACAGAAGCUGUUUCAGAAGAAACGAGGCCAGCAGAGAAACCAACAACUCCCUGACUUUCGGGCCCGAGGAAGCCCUCGCC 273 hIL12AB-AUGUGCCACCAGCAGCUGGUGAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCAGCCC 8TMCCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGUUGGAUUGGU NucleotideACCCCGACGCCCCCGGCGAGAUGGUGGUGCUGACCUGCGACACCCCCGAGGAGGAC SequenceGGCAUCACCUGGACCCUGGACCAGAGCAGCGAGGUGCUGGGCAGCGGCAAGACCCUGACCAUCCAGGUGAAGGAGUUCGGCGACGCCGGCCAGUACACCUGCCACAAGGGCGGCGAGGUGCUGAGCCACAGCCUGCUGCUGCUGCACAAGAAGGAGGACGGCAUCUGGAGCACCGACAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGAGAUGCGAGGCCAAGAACUACAGCGGCAGAUUCACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUGACCUUCAGCGUGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGUGACCUGCGGCGCCGCCACCCUGAGCGCCGAGAGAGUGAGAGGCGACAACAAGGAGUACGAGUACAGCGUGGAGUGCCAGGAAGAUAGCGCCUGCCCCGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUGCACAAGCUGAAGUACGAGAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAGCCCGACCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCCGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGAGCACCCCCCACAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGCAAGAGCAAGAGAGAGAAGAAAGAUAGAGUGUUCACCGACAAGACCAGCGCCACCGUGAUCUGCAGAAAGAACGCCAGCAUCAGCGUGAGAGCCCAAGAUAGAUACUACAGCAGCAGCUGGAGCGAGUGGGCCAGCGUGCCCUGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCUGCCCGUGGCCACCCCCGACCCCGGCAUGUUCCCCUGCCUGCACCACAGCCAGAACCUGCUGAGAGCCGUGAGCAACAUGCUGCAGAAGGCCCGGCAGACCCUGGAGUUCUACCCCUGCACCAGCGAGGAGAUCGACCACGAAGAUAUCACCAAAGAUAAGACCAGCACCGUGGAGGCCUGCCUGCCCCUGGAGCUGACCAAGAACGAGAGCUGCCUGAACAGCAGAGAGACCAGCUUCAUCACCAACGGCAGCUGCCUGGCCAGCAGAAAGACCAGCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGGCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAGACCGUGCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCAGAAUCAGAGCCGUGACCAUCGACAGAGUGAUGAGCUACCUGAACGCCAGCUCUGGUGGCGGAUCAGGCGGCGGCGGUUCAGGAGGCGGUGGAAGUGGAGGUGGCGGGUCUGGCGGAGGUUCACUGCAGAUCUACAUCUGGGCUCCACUGGCCGGCACCUGCGGCGUGCUGCUGCUGAGCCUGGUGAUCACCCUGUACUGCUACGGGAAACCAAUUCCAAAUCCCCUCCUGGGGUUGGAUAGCACC 274 hIL12AB-AUGUGCCACCAGCAGCUGGUGAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCAGCCC 8TM noCCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGUUGGAUUGGU epitope tagACCCCGACGCCCCCGGCGAGAUGGUGGUGCUGACCUGCGACACCCCCGAGGAGGAC NucleotideGGCAUCACCUGGACCCUGGACCAGAGCAGCGAGGUGCUGGGCAGCGGCAAGACCCU SequenceGACCAUCCAGGUGAAGGAGUUCGGCGACGCCGGCCAGUACACCUGCCACAAGGGCGGCGAGGUGCUGAGCCACAGCCUGCUGCUGCUGCACAAGAAGGAGGACGGCAUCUGGAGCACCGACAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGAGAUGCGAGGCCAAGAACUACAGCGGCAGAUUCACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUGACCUUCAGCGUGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGUGACCUGCGGCGCCGCCACCCUGAGCGCCGAGAGAGUGAGAGGCGACAACAAGGAGUACGAGUACAGCGUGGAGUGCCAGGAAGAUAGCGCCUGCCCCGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUGCACAAGCUGAAGUACGAGAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAGCCCGACCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCCGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGAGCACCCCCCACAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGCAAGAGCAAGAGAGAGAAGAAAGAUAGAGUGUUCACCGACAAGACCAGCGCCACCGUGAUCUGCAGAAAGAACGCCAGCAUCAGCGUGAGAGCCCAAGAUAGAUACUACAGCAGCAGCUGGAGCGAGUGGGCCAGCGUGCCCUGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCUGCCCGUGGCCACCCCCGACCCCGGCAUGUUCCCCUGCCUGCACCACAGCCAGAACCUGCUGAGAGCCGUGAGCAACAUGCUGCAGAAGGCCCGGCAGACCCUGGAGUUCUACCCCUGCACCAGCGAGGAGAUCGACCACGAAGAUAUCACCAAAGAUAAGACCAGCACCGUGGAGGCCUGCCUGCCCCUGGAGCUGACCAAGAACGAGAGCUGCCUGAACAGCAGAGAGACCAGCUUCAUCACCAACGGCAGCUGCCUGGCCAGCAGAAAGACCAGCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGGCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAGACCGUGCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCAGAAUCAGAGCCGUGACCAUCGACAGAGUGAUGAGCUACCUGAACGCCAGCUCUGGUGGCGGAUCAGGCGGCGGCGGUUCAGGAGGCGGUGGAAGUGGAGGUGGCGGGUCUGGCGGAGGUUCACUGCAGAUCUACAUCUGGGCUCCACUGGCCGGCACCUGCGGCGUGCUGCUGCUGAGCCUGGUGAUCAC CCUGUACUGCUAC275 h12AB-80TID AUGUGCCACCAGCAGCUGGUGAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCAGCCCNucleotide CCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGUUGGAUUGGUSequence 1 ACCCCGACGCCCCCGGCGAGAUGGUGGUGCUGACCUGCGACACCCCCGAGGAGGACmatched GGCAUCACCUGGACCCUGGACCAGAGCAGCGAGGUGCUGGGCAGCGGCAAGACCCUGACCAUCCAGGUGAAGGAGUUCGGCGACGCCGGCCAGUACACCUGCCACAAGGGCGGCGAGGUGCUGAGCCACAGCCUGCUGCUGCUGCACAAGAAGGAGGACGGCAUCUGGAGCACCGACAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGAGAUGCGAGGCCAAGAACUACAGCGGCAGAUUCACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUGACCUUCAGCGUGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGUGACCUGCGGCGCCGCCACCCUGAGCGCCGAGAGAGUGAGAGGCGACAACAAGGAGUACGAGUACAGCGUGGAGUGCCAGGAAGAUAGCGCCUGCCCCGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUGCACAAGCUGAAGUACGAGAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAGCCCGACCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCCGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGAGCACCCCCCACAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGCAAGAGCAAGAGAGAGAAGAAAGAUAGAGUGUUCACCGACAAGACCAGCGCCACCGUGAUCUGCAGAAAGAACGCCAGCAUCAGCGUGAGAGCCCAAGAUAGAUACUACAGCAGCAGCUGGAGCGAGUGGGCCAGCGUGCCCUGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCUGCCCGUGGCCACCCCCGACCCCGGCAUGUUCCCCUGCCUGCACCACAGCCAGAACCUGCUGAGAGCCGUGAGCAACAUGCUGCAGAAGGCCCGGCAGACCCUGGAGUUCUACCCCUGCACCAGCGAGGAGAUCGACCACGAAGAUAUCACCAAAGAUAAGACCAGCACCGUGGAGGCCUGCCUGCCCCUGGAGCUGACCAAGAACGAGAGCUGCCUGAACAGCAGAGAGACCAGCUUCAUCACCAACGGCAGCUGCCUGGCCAGCAGAAAGACCAGCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGGCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAGACCGUGCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCAGAAUCAGAGCCGUGACCAUCGACAGAGUGAUGAGCUACCUGAACGCCAGCUCUGGUGGCGGAUCAGGCGGCGGCGGUUCAGGAGGCGGUGGAAGUGGAGGUGGCGGGUCUGGCGGAGGUUCACUGCAGCUGCUGCCCAGCUGGGCCAUCACCCUGAUCAGCGUGAACGGCAUCUUCGUGAUCUGCUGCCUGACCUACUGCUUCGCCCCUCGAUGCAGAGAGAGAAGAAGAAACGAGAGACUGAGAAGAGAGAGCGUGCGACCCGUG 276 h12AB-80TIDAUGUGCCACCAGCAGCUGGUGAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCAGCCC NucleotideUCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGCUGGACUGGU Sequence 2ACCCUGACGCCCCUGGCGAGAUGGUGGUGCUGACCUGCGACACCCCUGAGGAGGAC SE_IL12_041GGCAUCACCUGGACCCUGGACCAGAGCAGCGAGGUGCUGGGCAGCGGCAAGACCCUGACCAUCCAGGUGAAGGAGUUCGGCGACGCCGGCCAGUACACCUGCCACAAGGGCGGAGAGGUGUUAAGCCACAGCCUGCUCUUGCUACACAAGAAGGAGGACGGUAUUUGGAGCACCGACAUCCUGAAGGACCAGAAGGAGCCUAAGAACAAGACCUUCCUGAGGUGCGAGGCCAAGAACUACAGCGGCAGAUUCACCUGCUGGUGGCUGACCACCAUCUCCACGGACCUGACCUUCAGCGUUAAGAGUAGCAGAGGCAGCAGCGACCCUCAGGGCGUGACUUGUGGCGCCGCCACCCUGAGCGCCGAGAGAGUGAGAGGCGACAACAAGGAGUACGAGUACAGCGUGGAGUGCCAGGAGGACAGCGCCUGCCCUGCCGCCGAGGAGAGCCUGCCUAUCGAGGUGAUGGUGGACGCCGUGCACAAGCUGAAGUACGAGAAUUACACCUCAUCCUUCUUCAUCAGAGACAUCAUCAAGCCUGACCCUCCAAAGAAUCUGCAGCUGAAGCCUCUGAAGAACAGCAGACAGGUGGAGGUGAGCUGGGAGUAUCCGGAUACCUGGAGCACACCUCACAGCUACUUCUCACUUACAUUCUGCGUGCAGGUGCAGGGCAAGAGCAAGAGAGAGAAGAAGGAUAGAGUGUUCACCGACAAGACCAGCGCCACCGUGAUCUGCAGAAAGAACGCCAGCAUCUCAGUGAGAGCCCAGGACAGAUACUACUCAUCCUCCUGGAGCGAGUGGGCCAGCGUGCCUUGCUCCGGUGGUGGUGGCGGAGGCAGCAGAAACCUGCCUGUGGCUACACCUGAUCCUGGCAUGUUCCCUUGCCUGCACCACAGCCAGAACCUGCUGAGAGCCGUGAGCAACAUGCUGCAGAAGGCCAGACAGACUCUGGAGUUCUACCCUUGCACCAGCGAGGAGAUCGACCACGAGGACAUCACCAAGGAUAAGACAAGCACCGUGGAGGCCUGCCUGCCUCUGGAGCUGACCAAGAACGAGAGCUGCCUAAACUCUAGGGAAACCAGCUUCAUUACUAACGGCAGUUGCUUAGCCAGCCGGAAGACAUCGUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAGAUGUACCAAGUGGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGACCCUAAGAGACAGAUCUUCCUAGACCAGAACAUGCUCGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAAACUGUGCCUCAGAAGAGUUCACUGGAGGAGCCUGACUUCUAUAAGACUAAGAUCAAGCUGUGUAUUCUCCUCCACGCCUUCAGAAUCAGGGCUGUCACCAUCGAUAGGGUGAUGAGCUACCUGAACGCAUCGUCCGGCGGAGGAUCCGGAGGAGGAGGCUCCGGCGGUGGUGGAAGUGGAGGAGGUGGAUCAGGAGGCGGUAGUCUCCAGCUCCUGCCUAGCUGGGCCAUCACCCUGAUCUCUGUAAACGGCAUUUUCGUCAUUUGCUGUCUGACUUACUGCUUCGCCCCUAGGUGCCGGGAGCGUAGGAGAAACGAGAGACUGCGCCGGGAGUCCGUGCGGCCUGUG 277 h12AB-80TIDAUGUGUCACCAGCAGCUCGUGAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCUCCCC NucleotideGCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGCUGGACUGGU Sequence 3ACCCCGACGCUCCCGGCGAGAUGGUGGUGCUGACCUGCGACACACCGGAGGAAGAC SE_IL12_042GGAAUCACCUGGACCCUGGACCAAUCCUCCGAAGUUCUGGGCAGCGGCAAGACCCUGACCAUCCAGGUGAAGGAGUUCGGCGACGCCGGCCAGUACACCUGCCACAAGGGCGGCGAGGUGCUGAGCCACAGCCUGCUCCUCCUCCACAAGAAAGAGGACGGCAUCUGGAGCACCGACAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGCGGUGCGAGGCCAAGAACUACUCAGGCCGAUUCACCUGUUGGUGGCUCACAACUAUCAGCACAGACCUGACCUUCAGCGUGAAGUCUAGCCGGGGCAGCAGCGAUCCUCAGGGCGUGACCUGCGGCGCCGCCACCCUGAGCGCCGAGCGGGUGCGGGGCGACAACAAGGAGUACGAGUACAGCGUGGAGUGCCAGGAGGACAGCGCCUGCCCGGCCGCCGAAGAGUCCCUGCCCAUCGAGGUGAUGGUGGACGCCGUGCACAAACUCAAGUACGAGAACUACACCUCCAGCUUCUUCAUCCGGGACAUCAUCAAGCCCGAUCCGCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCCGGCAGGUGGAGGUGAGCUGGGAAUACCCGGACACGUGGUCCACCCCACACAGCUACUUCAGCCUGACCUUUUGCGUGCAGGUCCAAGGCAAGAGCAAGCGGGAGAAGAAGGACCGGGUGUUCACCGAUAAGACCUCAGCCACCGUGAUUUGCAGAAAGAACGCAUCCAUAUCCGUACGCGCCCAGGAUCGGUACUACAGCAGCAGCUGGAGCGAGUGGGCCAGCGUGCCCUGUAGCGGCGGCGGCGGUGGUGGGAGUCGCAACCUGCCCGUGGCCACCCCGGACCCCGGCAUGUUCCCCUGCCUGCACCACAGCCAGAACCUGCUGCGGGCCGUGAGCAACAUGCUGCAGAAGGCCCGGCAGACCCUGGAGUUCUACCCCUGCACCAGCGAGGAGAUCGACCACGAGGACAUCACCAAGGACAAGACCAGCACCGUGGAGGCCUGCCUGCCCCUGGAGCUGACCAAGAACGAGAGCUGCCUGAACAGUCGCGAAACCUCCUUCAUUACGAACGGCAGCUGCCUGGCCAGCCGGAAGACCAGCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGGCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAAACCGUGCCCCAGAAGUCCAGCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCCGGAUCCGGGCCGUGACCAUCGACCGGGUGAUGAGCUACCUGAACGCCUCUUCCGGUGGCGGGAGCGGAGGCGGUGGAUCUGGCGGAGGAGGGUCGGGAGGCGGCGGAAGCGGUGGUGGAAGCCUUCAACUGCUGCCCUCGUGGGCCAUCACACUGAUCUCCGUGAACGGCAUCUUCGUGAUCUGCUGCCUGACCUACUGCUUCGCCCCUCGGUGCCGCGAGCGACGGAGAAACGAGAGGCUCAGACGGGAGAGCGUGCGGCCCGUG 278 h12AB-80TIDAUGUGCCACCAACAGCUGGUGAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCUCACC NucleotideCCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGCUGGACUGGU Sequence 4ACCCCGACGCCCCUGGCGAGAUGGUGGUGCUGACCUGCGACACGCCCGAGGAAGAC SE_IL12_043GGUAUCACCUGGACUCUGGACCAGAGCAGCGAGGUGCUGGGCAGCGGCAAGACCCUGACCAUCCAGGUGAAGGAGUUCGGCGACGCCGGCCAGUACACCUGCCACAAGGGCGGCGAAGUGCUGAGCCACAGCCUUCUGCUGCUGCACAAGAAGGAGGACGGCAUUUGGAGCACCGACAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGCGGUGCGAGGCCAAGAACUACAGCGGCCGGUUCACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUUACCUUCAGCGUUAAGAGCAGCCGGGGCAGCAGCGAUCCCCAGGGCGUGACCUGCGGAGCCGCCACCCUCUCCGCAGAGCGGGUGCGUGGCGACAACAAGGAGUACGAGUACAGCGUGGAGUGCCAGGAGGAUAGCGCCUGUCCCGCUGCCGAAGAGAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUGCACAAGCUGAAGUACGAGAACUACACCAGCAGCUUCUUCAUCCGGGACAUCAUCAAGCCCGAUCCACCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCAGGCAGGUUGAGGUGAGCUGGGAAUACCCCGACACCUGGAGCACCCCUCACAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGCAAGAGCAAGCGGGAGAAGAAGGAUCGGGUGUUCACCGAUAAGACCAGCGCCACCGUGAUCUGCCGGAAGAACGCCAGCAUCAGCGUUCGGGCCCAGGACCGGUACUACAGCAGCAGCUGGAGCGAGUGGGCCAGCGUGCCCUGCUCUGGAGGCGGAGGCGGAGGCUCACGGAACCUGCCAGUGGCCACGCCGGAUCCCGGCAUGUUCCCCUGCCUGCACCACAGCCAGAACCUGCUGCGGGCCGUGAGCAACAUGCUGCAGAAGGCCCGGCAGACCCUGGAGUUCUACCCCUGCACCAGCGAGGAGAUCGACCACGAGGACAUCACCAAGGACAAGACCAGCACCGUGGAGGCCUGCCUGCCCCUGGAGCUGACCAAGAACGAGAGCUGCCUGAACAGCCGGGAGACAAGCUUCAUCACCAACGGCAGCUGCCUGGCCAGCCGGAAGACCAGCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGGCAGAUCUUCCUGGAUCAGAACAUGCUGGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAGACUGUGCCCCAGAAGUCCAGCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCCGGAUCCGGGCUGUGACCAUCGACCGGGUGAUGAGCUACCUGAACGCCUCUUCCGGCGGCGGAUCGGGAGGUGGAGGUUCUGGAGGAGGUGGAAGCGGUGGUGGCGGAAGCGGCGGUGGCAGCCUGCAAUUGCUCCCCAGCUGGGCCAUCACCCUGAUCAGCGUGAACGGCAUCUUCGUGAUCUGUUGCCUGACCUACUGCUUCGCCCCACGGUGCCGGGAGAGACGGCGGAACGAGCGGCUGCGGCGAGAGAGCGUGCGGCCCGUG 279 h12AB-80TIDAUGUGUCACCAGCAGUUGGUGAUAUCUUGGUUCUCACUGGUGUUUCUUGCAUCACC NucleotideACUCGUGGCGAUCUGGGAACUUAAGAAGGACGUCUACGUGGUGGAGUUAGAUUGGU Sequence 5AUCCUGACGCACCCGGGGAAAUGGUUGUCCUCACGUGCGACACUCCAGAGGAAGAC SE_IL12_044GGGAUCACCUGGACCCUGGAUCAGUCGUCAGAGGUACUUGGCAGUGGCAAGACACUGACAAUCCAGGUUAAAGAGUUUGGUGACGCCGGGCAGUAUACGUGCCACAAGGGCGGCGAGGUGUUGUCACAUUCUCUGCUUCUCCUGCACAAGAAAGAAGACGGCAUCUGGUCAACUGACAUCCUGAAAGACCAGAAAGAACCCAAGAAUAAGACCUUCCUCCGUUGCGAAGCAAAGAACUACUCAGGGCGUUUCACUUGCUGGUGGCUAACCACAAUUUCUACCGAUCUGACGUUCUCUGUGAAGUCAAGUAGGGGAUCCUCAGACCCUCAAGGGGUCACCUGCGGCGCCGCCACCUUAUCCGCCGAAAGAGUUCGGGGUGACAAUAAAGAGUACGAGUACAGCGUCGAGUGUCAGGAGGACUCCGCCUGUCCUGCUGCAGAGGAGUCCCUGCCGAUCGAAGUUAUGGUGGACGCCGUCCACAAGCUCAAAUACGAGAACUACACCUCAAGCUUCUUCAUCAGAGACAUCAUCAAGCCUGAUCCACCCAAGAACCUGCAACUGAAGCCUUUGAAGAACAGCCGACAGGUGGAGGUUUCUUGGGAAUAUCCAGACACGUGGAGUACGCCCCAUUCCUACUUCAGCUUGACCUUCUGCGUGCAGGUUCAGGGGAAGUCCAAGAGAGAGAAGAAGGAUCGUGUGUUCACAGACAAGACCUCCGCCACCGUGAUCUGCCGGAAGAACGCAUCUAUCAGUGUUAGGGCCCAGGAUCGGUACUACUCGAGUUCCUGGUCUGAGUGGGCAAGUGUGCCCUGCUCCGGUGGCGGCGGAGGAGGGUCAAGGAACCUGCCCGUUGCCACACCAGAUCCAGGAAUGUUCCCCUGUCUGCACCACUCUCAGAACCUUUUGCGAGCCGUUUCUAAUAUGCUUCAGAAGGCUCGGCAGACCCUUGAGUUUUAUCCCUGCACGUCUGAGGAGAUCGAUCACGAGGACAUCACCAAGGACAAGACUUCCACCGUUGAAGCCUGUUUACCUCUGGAACUGACCAAGAACGAAUCCUGUCUCAACAGUAGGGAAACGAGCUUCAUCACUAACGGAAGCUGUCUGGCUAGCCGGAAGACCUCUUUUAUGAUGGCCCUGUGCUUGAGCUCUAUUUACGAAGAUUUGAAGAUGUACCAAGUGGAAUUUAAGACUAUGAACGCCAAACUGCUGAUGGACCCUAAGCGCCAAAUCUUCUUGGAUCAGAAUAUGCUGGCUGUAAUCGACGAGCUCAUGCAGGCUCUGAACUUCAACAGCGAGACGGUACCGCAGAAGAGUUCCCUGGAAGAACCGGACUUCUACAAGACUAAGAUUAAACUCUGCAUACUCCUCCACGCCUUCCGGAUCAGGGCCGUCACAAUAGAUAGGGUCAUGAGUUAUCUUAACGCGAGUUCUGGUGGUGGAUCGGGUGGCGGAGGCUCAGGAGGAGGCGGUUCUGGCGGUGGUGGGAGUGGAGGCGGUAGUCUGCAGCUGCUGCCGAGUUGGGCAAUCACGCUAAUCAGCGUGAACGGAAUAUUCGUAAUUUGUUGCCUCACCUAUUGUUUCGCACCCAGGUGCAGGGAAAGGAGGCGAAACGAAAGGUUGAGGAGGGAAUCUGUCCGGCCAGUG 280 HumanCAGAAGAAGCCCAGAUACGAGAUCCGGUGGAAGGUGAUCGAGAGCGUGAGCAGCGA PGFRBCGGCCACGAGUUCAUCUUCGUGGACCCCAUGCAGCUGCCCUACGACAGCACCUGGG G739trAGCUGCCCCGUGAUCAGCUGGUGCUGGGCAGAACCCUGGGCAGCGGCGCCUUCGGC intracellularCAGGUGGUGGAGGCUACCGCCCACGGCCUGAGCCACAGCCAGGCCACCAUGAAGGU domainGGCCGUGGCCAUGCUCAAGAGCACCGCCAGAAGCAGCGAGAAGCAGGCCCUGAUGA nucleotideGCGAGCUGAAGAUCAUGAGCCAUCUGGGGCCCCACCUGAACGUGGUGAACCUGCUG sequenceGGCGCCUGCACCAAGGGCGGCCCCAUCUACAUCAUCACCGAGUACUGCAGAUACGGCGACCUGGUGGACUACCUGCACAGAAACAAGCACACCUUCCUGCAGCACCACAGCGACAAGAGAAGACCUCCCAGCGCCGAGCUGUACAGCAACGCCCUGCCCGUUGGUCUGCCCCUACCCAGCCACGUGAGCCUGACCGGCGAGAGCGACGGCGGC 281 h12AB-PTM-AUGUGCCACCAGCAGCUGGUGAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCAGCCC ICD-E570trCCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGUUGGAUUGGU NucleotideACCCCGACGCCCCCGGCGAGAUGGUGGUGCUGACCUGCGACACCCCCGAGGAGGAC SequenceGGCAUCACCUGGACCCUGGACCAGAGCAGCGAGGUGCUGGGCAGCGGCAAGACCCUGACCAUCCAGGUGAAGGAGUUCGGCGACGCCGGCCAGUACACCUGCCACAAGGGCGGCGAGGUGCUGAGCCACAGCCUGCUGCUGCUGCACAAGAAGGAGGACGGCAUCUGGAGCACCGACAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGAGAUGCGAGGCCAAGAACUACAGCGGCAGAUUCACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUGACCUUCAGCGUGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGUGACCUGCGGCGCCGCCACCCUGAGCGCCGAGAGAGUGAGAGGCGACAACAAGGAGUACGAGUACAGCGUGGAGUGCCAGGAAGAUAGCGCCUGCCCCGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUGCACAAGCUGAAGUACGAGAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAGCCCGACCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCCGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGAGCACCCCCCACAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGCAAGAGCAAGAGAGAGAAGAAAGAUAGAGUGUUCACCGACAAGACCAGCGCCACCGUGAUCUGCAGAAAGAACGCCAGCAUCAGCGUGAGAGCCCAAGAUAGAUACUACAGCAGCAGCUGGAGCGAGUGGGCCAGCGUGCCCUGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCUGCCCGUGGCCACCCCCGACCCCGGCAUGUUCCCCUGCCUGCACCACAGCCAGAACCUGCUGAGAGCCGUGAGCAACAUGCUGCAGAAGGCCCGGCAGACCCUGGAGUUCUACCCCUGCACCAGCGAGGAGAUCGACCACGAAGAUAUCACCAAAGAUAAGACCAGCACCGUGGAGGCCUGCCUGCCCCUGGAGCUGACCAAGAACGAGAGCUGCCUGAACAGCAGAGAGACCAGCUUCAUCACCAACGGCAGCUGCCUGGCCAGCAGAAAGACCAGCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGGCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAGACCGUGCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCAGAAUCAGAGCCGUGACCAUCGACAGAGUGAUGAGCUACCUGAACGCCAGCUCUGGUGGCGGAUCAGGCGGCGGCGGUUCAGGAGGCGGUGGAAGUGGAGGUGGCGGGUCUGGCGGAGGUUCACUGCAGGUGGUGGUGAUCAGCGCCAUCCUGGCCCUGGUGGUGCUCACCAUCAUCAGCCUGAUCAUCCUGAUCAUGCUGUGGCAGAAGAAGCCCAGAUACGAGAUCCGGUGGAAGGUGAUCGAGAGCGUGAGCAGCGACGGCCACGAGUUCAUCUUCGUGGACCCCAUGCAGCUGCCCUACGACAGCACCUGGGAGCUGCCCCGUGAUCAGCUGGUGCUGGGCAGAACCCUGGGCAGCGGCGCCUUCGGCCAGGUGGUGGAGGCUACCGCCCACGGCCUGAGCCACAGCCAGGCCACCAUGAAGGUGGCCGUGGCCAUGCUCAAGAGCACCGCCAGAAGCAGCGAGAAGCAGGCCCUGAUGAGCGAGCUGAAGAUCAUGAGCCAUCUGGGGCCCCACCUGAACGUGGUGAACCUGCUGGGCGCCUGCACCAAGGGCGGCCCCAUCUACAUCAUCACCGAGUACUGCAGAUACGGCGACCUGGUGGACUACCUGCACAGAAACAAGCACACCUUCCUGCAGCACCACAGCGACAAGAGAAGACCUCCCAGCGCCGAGCUGUACAGCAACGCCCUGCCCGUUGGUCUGCCCCUACCCAGCCACGUGAGCCUGACCGGCGAGAGCGACGGCG GC 282h12AB-PTM- AUGUGCCACCAGCAGCUGGUGAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCAGCCCICD-G739tr CCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGUUGGAUUGGUNucleotide ACCCCGACGCCCCCGGCGAGAUGGUGGUGCUGACCUGCGACACCCCCGAGGAGGACSequence GGCAUCACCUGGACCCUGGACCAGAGCAGCGAGGUGCUGGGCAGCGGCAAGACCCUGACCAUCCAGGUGAAGGAGUUCGGCGACGCCGGCCAGUACACCUGCCACAAGGGCGGCGAGGUGCUGAGCCACAGCCUGCUGCUGCUGCACAAGAAGGAGGACGGCAUCUGGAGCACCGACAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGAGAUGCGAGGCCAAGAACUACAGCGGCAGAUUCACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUGACCUUCAGCGUGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGUGACCUGCGGCGCCGCCACCCUGAGCGCCGAGAGAGUGAGAGGCGACAACAAGGAGUACGAGUACAGCGUGGAGUGCCAGGAAGAUAGCGCCUGCCCCGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUGCACAAGCUGAAGUACGAGAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAGCCCGACCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCCGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGAGCACCCCCCACAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGCAAGAGCAAGAGAGAGAAGAAAGAUAGAGUGUUCACCGACAAGACCAGCGCCACCGUGAUCUGCAGAAAGAACGCCAGCAUCAGCGUGAGAGCCCAAGAUAGAUACUACAGCAGCAGCUGGAGCGAGUGGGCCAGCGUGCCCUGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCUGCCCGUGGCCACCCCCGACCCCGGCAUGUUCCCCUGCCUGCACCACAGCCAGAACCUGCUGAGAGCCGUGAGCAACAUGCUGCAGAAGGCCCGGCAGACCCUGGAGUUCUACCCCUGCACCAGCGAGGAGAUCGACCACGAAGAUAUCACCAAAGAUAAGACCAGCACCGUGGAGGCCUGCCUGCCCCUGGAGCUGACCAAGAACGAGAGCUGCCUGAACAGCAGAGAGACCAGCUUCAUCACCAACGGCAGCUGCCUGGCCAGCAGAAAGACCAGCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGGCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAGACCGUGCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCAGAAUCAGAGCCGUGACCAUCGACAGAGUGAUGAGCUACCUGAACGCCAGCUCUGGUGGCGGAUCAGGCGGCGGCGGUUCAGGAGGCGGUGGAAGUGGAGGUGGCGGGUCUGGCGGAGGUUCACUGCAGGUGGUGGUGAUCAGCGCCAUCCUGGCCCUGGUGGUGCUCACCAUCAUCAGCCUGAUCAUCCUGAUCAUGCUGUGGCAGAAGAAGCCCAGAUACGAGAUCCGGUGGAAGGUGAUCGAGAGCGUGAGCAGCGACGGCCACGAGUUCAUCUUCGUGGACCCCAUGCAGCUGCCCUACGACAGCACCUGGGAGCUGCCCCGUGAUCAGCUGGUGCUGGGCAGAACCCUGGGCAGCGGCGCCUUCGGCCAGGUGGUGGAGGCUACCGCCCACGGCCUGAGCCACAGCCAGGCCACCAUGAAGGUGGCCGUGGCCAUGCUCAAGAGCACCGCCAGAAGCAGCGAGAAGCAGGCCCUGAUGAGCGAGCUGAAGAUCAUGAGCCAUCUGGGGCCCCACCUGAACGUGGUGAACCUGCUGGGCGCCUGCACCAAGGGCGGCCCCAUCUACAUCAUCACCGAGUACUGCAGAUACGGCGACCUGGUGGACUACCUGCACAGAAACAAGCACACCUUCCUGCAGCACCACAGCGACAAGAGAAGACCUCCCAGCGCCGAGCUGUACAGCAACGCCCUGCCCGUUGGUCUGCCCCUACCCAGCCACGUGAGCCUGACCGGCGAGAGCGACGGCG GC 2833′UTR with UGAUAAUAGGCUGGAGCCUCGGUGGCCUAGCUUCUUGCCCCUUGGGCCUCCCCCCAmir-122-5p GCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUbinding site GGUCUUUGAAUAAAGUCUGAGUGGGCGGC 284 Wild TypeAUAUGGGAACUGAAGAAAGAUGUUUAUGUCGUAGAAUUGGAUUGGUAUCCGGAUGC IL12B withoutCCCUGGAGAAAUGGUGGUCCUCACCUGUGACACCCCUGAAGAAGAUGGUAUCACCU signal (IL12B)GGACCUUGGACCAGAGCAGUGAGGUCUUAGGCUCUGGCAAAACCCUGACCAUCCAA Nucleic AcidsGUCAAAGAGUUUGGAGAUGCUGGCCAGUACACCUGUCACAAAGGAGGCGAGGUUCUAAGCCAUUCGCUCCUGCUGCUUCACAAAAAGGAAGAUGGAAUUUGGUCCACUGAUAUUUUAAAGGACCAGAAAGAACCCAAAAAUAAGACCUUUCUAAGAUGCGAGGCCAAGAAUUAUUCUGGACGUUUCACCUGCUGGUGGCUGACGACAAUCAGUACUGAUUUGACAUUCAGUGUCAAAAGCAGCAGAGGCUCUUCUGACCCCCAAGGGGUGACGUGCGGAGCUGCUACACUCUCUGCAGAGAGAGUCAGAGGGGACAACAAGGAGUAUGAGUACUCAGUGGAGUGCCAGGAGGACAGUGCCUGCCCAGCUGCUGAGGAGAGUCUGCCCAUUGAGGUCAUGGUGGAUGCCGUUCACAAGCUCAAGUAUGAAAACUACACCAGCAGCUUCUUCAUCAGGGACAUCAUCAAACCUGACCCACCCAAGAACUUGCAGCUGAAGCCAUUAAAGAAUUCUCGGCAGGUGGAGGUCAGCUGGGAGUACCCUGACACCUGGAGUACUCCACAUUCCUACUUCUCCCUGACAUUCUGCGUUCAGGUCCAGGGCAAGAGCAAGAGAGAAAAGAAAGAUAGAGUCUUCACGGACAAGACCUCAGCCACGGUCAUCUGCCGCAAAAAUGCCAGCAUUAGCGUGCGGGCCCAGGACCGCUACUAUAGCUCAUCUUGGAGCGAAUGGGCAUCUGUGCCCUGCAGU 285 Wild TypeAGAAACCUCCCCGUGGCCACUCCAGACCCAGGAAUGUUCCCAUGCCUUCACCACUC IL12A withoutCCAAAACCUGCUGAGGGCCGUCAGCAACAUGCUCCAGAAGGCCAGACAAACUCUAG signal peptideAAUUUUACCCUUGCACUUCUGAAGAGAUUGAUCAUGAAGAUAUCACAAAAGAUAAA Nucleic acidsACCAGCACAGUGGAGGCCUGUUUACCAUUGGAAUUAACCAAGAAUGAGAGUUGCCUAAAUUCCAGAGAGACCUCUUUCAUAACUAAUGGGAGUUGCCUGGCCUCCAGAAAGACCUCUUUUAUGAUGGCCCUGUGCCUUAGUAGUAUUUAUGAAGACUUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAAUGCAAAGCUUCUGAUGGAUCCUAAGAGGCAGAUCUUUCUAGAUCAAAACAUGCUGGCAGUUAUUGAUGAGCUGAUGCAGGCCCUGAAUUUCAACAGUGAGACUGUGCCACAAAAAUCCUCCCUUGAAGAACCGGAUUUUUAUAAAACUAAAAUCAAGCUCUGCAUACUUCUUCAUGCUUUCAGAAUUCGGGCAGUGACUAUUGAUAGAGUGAUGAGCUAUCUGAAUGCUUCC 286 5′UTRGGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGA 287 V1-5′UTRGGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGACCCCGGCGCCGCCACC 288 V2-5′UTRGGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGACCCCGGCGCCACC 289 StandardGGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACC 5′UTR 290 hIL12AB_001AUGUGUCACCAGCAGCUGGUCAUUAGCUGGUUUAGCCUUGUGUUCCUGGCCUCCCC ORFCCUUGUCGCUAUUUGGGAGCUCAAGAAGGACGUGUACGUGGUGGAGCUGGACUGGUACCCAGACGCGCCCGGAGAGAUGGUAGUUCUGACCUGUGAUACCCCAGAGGAGGACGGCAUCACCUGGACUCUGGACCAAAGCAGCGAGGUUUUGGGCUCAGGGAAAACGCUGACCAUCCAGGUGAAGGAAUUCGGCGACGCCGGACAGUACACCUGCCAUAAGGGAGGAGAGGUGCUGAGCCAUUCCCUUCUUCUGCUGCACAAGAAAGAGGACGGCAUCUGGUCUACCGACAUCCUGAAAGACCAGAAGGAGCCCAAGAACAAAACCUUCCUGAGGUGCGAGGCCAAGAACUACUCCGGCAGGUUCACUUGUUGGUGGCUGACCACCAUCAGUACAGACCUGACUUUUAGUGUAAAAAGCUCCAGAGGCUCGUCCGAUCCCCAAGGGGUGACCUGCGGCGCAGCCACUCUGAGCGCUGAGCGCGUGCGCGGUGACAAUAAAGAGUACGAGUACAGCGUUGAGUGUCAAGAAGACAGCGCUUGCCCUGCCGCCGAGGAGAGCCUGCCUAUCGAGGUGAUGGUUGACGCAGUGCACAAGCUUAAGUACGAGAAUUACACCAGCUCAUUCUUCAUUAGAGAUAUAAUCAAGCCUGACCCACCCAAGAACCUGCAGCUGAAGCCACUGAAAAACUCACGGCAGGUCGAAGUGAGCUGGGAGUACCCCGACACCUGGAGCACUCCUCAUUCCUAUUUCUCUCUUACAUUCUGCGUCCAGGUGCAGGGCAAGAGCAAGCGGGAAAAGAAGGAUCGAGUCUUCACCGACAAAACAAGCGCGACCGUGAUUUGCAGGAAGAACGCCAGCAUCUCCGUCAGAGCCCAGGAUAGAUACUAUAGUAGCAGCUGGAGCGAGUGGGCAAGCGUGCCCUGUUCCGGCGGCGGGGGCGGGGGCAGCCGAAACUUGCCUGUCGCUACCCCGGACCCUGGAAUGUUUCCGUGUCUGCACCACAGCCAGAACCUGCUGAGAGCCGUGUCGAAUAUGCUCCAGAAGGCCCGGCAGACCCUUGAGUUCUACCCCUGUACCAGCGAAGAGAUCGAUCAUGAGGACAUCACGAAAGACAAGACUUCCACCGUCGAGGCUUGUCUCCCGCUGGAGCUGACCAAGAACGAGAGCUGUCUGAAUAGCCGGGAGACAUCUUUCAUCACGAAUGGUAGCUGUCUGGCCAGCAGGAAAACUUCCUUCAUGAUGGCUCUCUGCCUGAGCUCUAUCUAUGAAGAUCUGAAGAUGUAUCAGGUGGAGUUUAAGACUAUGAACGCCAAACUCCUGAUGGACCCAAAAAGGCAAAUCUUUCUGGACCAGAAUAUGCUGGCCGUGAUAGACGAGCUGAUGCAGGCACUGAACUUCAACAGCGAGACAGUGCCACAGAAAUCCAGCCUGGAGGAGCCUGACUUUUACAAAACUAAGAUCAAGCUGUGUAUCCUGCUGCACGCCUUUAGAAUCCGUGCCGUGACUAUCGACAGGGUGAUGUCAUACCUCAACGCUUCA 291 hIL12AB_002AUGUGCCACCAGCAGCUGGUGAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCAGCCC ORFCCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGCUGGACUGGUACCCCGACGCCCCCGGCGAGAUGGUGGUGCUGACCUGCGACACCCCCGAGGAGGACGGCAUCACCUGGACCCUGGACCAGAGCAGCGAGGUGCUGGGCAGCGGCAAGACCCUGACCAUCCAGGUGAAGGAGUUCGGCGACGCCGGCCAGUACACCUGCCACAAGGGCGGCGAGGUGCUGAGCCACAGCCUGCUGCUGCUGCACAAGAAGGAGGACGGCAUCUGGAGCACCGACAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGAGAUGCGAGGCCAAGAACUACAGCGGCAGAUUCACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUGACCUUCAGCGUGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGUGACCUGCGGCGCCGCCACCCUGAGCGCCGAGAGAGUGAGAGGCGACAACAAGGAGUACGAGUACAGCGUGGAGUGCCAGGAGGACAGCGCCUGCCCCGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUGCACAAGCUGAAGUACGAGAACUACACCAGCAGCUUCUUCAUCAGAGACAUCAUCAAGCCCGACCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCAGACAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGAGCACCCCCCACAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGCAAGAGCAAGAGAGAGAAGAAGGACAGAGUGUUCACCGACAAGACCAGCGCCACCGUGAUCUGCAGAAAGAACGCCAGCAUCAGCGUGAGAGCCCAGGACAGAUACUACAGCAGCAGCUGGAGCGAGUGGGCCAGCGUGCCCUGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCUGCCCGUGGCCACCCCCGACCCCGGCAUGUUCCCCUGCCUGCACCACAGCCAGAACCUGCUGAGAGCCGUGAGCAACAUGCUGCAGAAGGCCAGACAGACCCUGGAGUUCUACCCCUGCACCAGCGAGGAGAUCGACCACGAGGACAUCACCAAGGACAAGACCAGCACCGUGGAGGCCUGCCUGCCCCUGGAGCUGACCAAGAACGAGAGCUGCCUGAACAGCAGAGAGACCAGCUUCAUCACCAACGGCAGCUGCCUGGCCAGCAGAAAGACCAGCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGAGACAGAUCUUCCUGGACCAGAACAUGCUGGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAGACCGUGCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCAGAAUCAGAGCCGUGACCAUCGACAGAGUGAUGAGCUACCUGAACGCCAGC 292 hIL12AB_003AUGUGUCACCAGCAGUUGGUCAUCUCUUGGUUUUCCCUGGUUUUUCUGGCAUCUCC ORFCCUCGUGGCCAUAUGGGAACUGAAGAAAGAUGUUUAUGUCGUAGAAUUGGAUUGGUAUCCGGAUGCCCCUGGAGAAAUGGUGGUCCUCACCUGUGACACCCCUGAAGAAGAUGGUAUCACCUGGACCUUGGACCAGAGCAGUGAGGUCUUAGGCUCUGGCAAAACCCUGACCAUCCAAGUCAAAGAGUUUGGAGAUGCUGGCCAGUACACCUGUCACAAAGGAGGCGAGGUUCUAAGCCAUUCGCUCCUGCUGCUUCACAAAAAGGAAGAUGGAAUUUGGUCCACUGAUAUUUUAAAGGACCAGAAAGAACCCAAAAAUAAGACCUUUCUAAGAUGCGAGGCCAAGAAUUAUUCUGGACGUUUCACCUGCUGGUGGCUGACGACAAUCAGUACUGAUUUGACAUUCAGUGUCAAAAGCAGCAGAGGCUCUUCUGACCCCCAAGGGGUGACGUGCGGAGCUGCUACACUCUCUGCAGAGAGAGUCAGAGGGGACAACAAGGAGUAUGAGUACUCAGUGGAGUGCCAGGAGGACAGUGCCUGCCCAGCUGCUGAGGAGAGUCUGCCCAUUGAGGUCAUGGUGGAUGCCGUUCACAAGCUCAAGUAUGAAAACUACACCAGCAGCUUCUUCAUCAGGGACAUCAUCAAACCUGACCCACCCAAGAACUUGCAGCUGAAGCCAUUAAAGAAUUCUCGGCAGGUGGAGGUCAGCUGGGAGUACCCUGACACCUGGAGUACUCCACAUUCCUACUUCUCCCUGACAUUCUGCGUUCAGGUCCAGGGCAAGAGCAAGAGAGAAAAGAAAGAUAGAGUCUUCACGGACAAGACCUCAGCCACGGUCAUCUGCCGCAAAAAUGCCAGCAUUAGCGUGCGGGCCCAGGACCGCUACUAUAGCUCAUCUUGGAGCGAAUGGGCAUCUGUGCCCUGCAGUGGCGGAGGGGGCGGAGGGAGCAGAAACCUCCCCGUGGCCACUCCAGACCCAGGAAUGUUCCCAUGCCUUCACCACUCCCAAAACCUGCUGAGGGCCGUCAGCAACAUGCUCCAGAAGGCCAGACAAACUUUAGAAUUUUACCCUUGCACUUCUGAAGAGAUUGAUCAUGAAGAUAUCACAAAAGAUAAAACCAGCACAGUGGAGGCCUGUUUACCAUUGGAAUUAACCAAGAAUGAGAGUUGCCUAAAUUCCAGAGAGACCUCUUUCAUAACUAAUGGGAGUUGCCUGGCCUCCAGAAAGACCUCUUUUAUGAUGGCCCUGUGCCUUAGUAGUAUUUAUGAAGACUUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAAUGCAAAGCUUCUGAUGGAUCCUAAGAGGCAGAUCUUUUUAGAUCAAAACAUGCUGGCAGUUAUUGAUGAGCUGAUGCAGGCCCUGAAUUUCAACAGUGAGACUGUGCCACAAAAAUCCUCCCUUGAAGAACCGGACUUCUACAAGACCAAGAUCAAGCUCUGCAUACUUCUUCAUGCUUUCAGAAUUCGGGCAGUGACUAUUGAUAGAGUGAUGAGCUAUCUGAAUGCUUCC 293 hIL12AB_004AUGGGCUGCCACCAGCAGCUGGUCAUCAGCUGGUUCUCCCUGGUCUUCCUGGCCAG ORFCCCCCUGGUGGCCAUCUGGGAGCUGAAGAAAGAUGUCUAUGUUGUAGAGCUGGACUGGUACCCAGAUGCUCCUGGAGAAAUGGUGGUUCUCACCUGUGACACGCCAGAAGAAGAUGGCAUCACCUGGACGCUGGACCAGAGCUCAGAAGUUCUUGGCAGUGGAAAAACGCUGACCAUACAAGUAAAAGAAUUUGGGGAUGCUGGCCAGUACACCUGCCACAAAGGAGGAGAAGUUCUCAGCCACAGCCUGCUGCUGCUGCACAAGAAAGAAGAUGGCAUCUGGAGCACAGACAUUUUAAAAGACCAGAAGGAGCCCAAGAACAAAACCUUCCUUCGAUGUGAGGCCAAGAACUACAGUGGCCGCUUCACCUGCUGGUGGCUCACCACCAUCAGCACAGACCUCACCUUCUCGGUGAAGAGCAGCCGUGGCAGCUCAGACCCCCAAGGAGUCACCUGUGGGGCGGCCACGCUGUCGGCAGAAAGAGUUCGAGGGGACAACAAGGAAUAUGAAUACUCGGUGGAAUGUCAAGAAGACUCGGCCUGCCCGGCGGCAGAAGAAAGUCUUCCCAUAGAAGUCAUGGUGGAUGCUGUUCACAAAUUAAAAUAUGAAAACUACACCAGCAGCUUCUUCAUCAGAGACAUCAUCAAGCCAGACCCGCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCAGACAAGUGGAAGUUUCCUGGGAGUACCCAGACACGUGGAGCACGCCGCACAGCUACUUCAGCCUCACCUUCUGUGUACAAGUACAAGGCAAGAGCAAGAGAGAGAAGAAAGAUCGUGUCUUCACAGACAAAACCUCGGCGACGGUCAUCUGCAGGAAGAAUGCCUCCAUCUCGGUUCGAGCCCAGGACCGCUACUACAGCAGCAGCUGGAGUGAGUGGGCCUCGGUGCCCUGCAGUGGUGGCGGCGGCGGCGGCAGCAGAAACCUUCCUGUGGCCACGCCGGACCCUGGCAUGUUCCCGUGCCUGCACCACAGCCAAAAUUUACUUCGAGCUGUUUCUAACAUGCUGCAGAAAGCAAGACAAACUUUAGAAUUCUACCCCUGCACCUCAGAAGAAAUAGACCAUGAAGACAUCACCAAAGAUAAAACCAGCACUGUAGAGGCCUGCCUGCCCCUGGAGCUCACCAAGAAUGAAUCCUGCCUCAACAGCAGAGAGACCAGCUUCAUCACCAAUGGCAGCUGCCUGGCCAGCAGGAAAACCAGCUUCAUGAUGGCGCUCUGCCUGAGCAGCAUCUAUGAAGAUUUGAAGAUGUACCAAGUAGAAUUUAAAACCAUGAAUGCCAAGCUGCUCAUGGACCCCAAGAGACAAAUAUUUUUGGAUCAAAACAUGCUGGCUGUCAUUGAUGAGCUCAUGCAAGCAUUAAACUUCAACUCAGAGACGGUGCCCCAGAAGAGCAGCCUGGAGGAGCCAGACUUCUACAAAACCAAGAUCAAGCUCUGCAUCUUAUUACAUGCCUUCCGCAUCCGGGCGGUCACCAUUGACCGUGUCAUGUCCUACUUAAAUGCCAGC 294 hIL12AB_005AUGUGCCACCAGCAGCUGGUCAUCAGCUGGUUCUCCCUGGUCUUCCUGGCCAGCCC ORFCCUGGUGGCCAUCUGGGAGCUGAAGAAAGAUGUCUAUGUUGUAGAGCUGGACUGGUACCCAGAUGCUCCUGGAGAAAUGGUGGUUCUCACCUGUGACACGCCAGAAGAAGAUGGCAUCACCUGGACGCUGGACCAGAGCUCAGAAGUUCUUGGCAGUGGAAAAACGCUGACCAUACAAGUAAAAGAAUUUGGGGAUGCUGGCCAGUACACCUGCCACAAAGGAGGAGAAGUUCUCAGCCACAGCCUGCUGCUGCUGCACAAGAAAGAAGAUGGCAUCUGGAGCACAGACAUUUUAAAAGACCAGAAGGAGCCCAAGAACAAAACCUUCCUUCGAUGUGAGGCCAAGAACUACAGUGGCCGCUUCACCUGCUGGUGGCUCACCACCAUCAGCACAGACCUCACCUUCUCGGUGAAGAGCAGCCGUGGCAGCUCAGACCCCCAAGGAGUCACCUGUGGGGCGGCCACGCUGUCGGCAGAAAGAGUUCGAGGGGACAACAAGGAAUAUGAAUACUCGGUGGAAUGUCAAGAAGACUCGGCCUGCCCGGCGGCAGAAGAAAGUCUUCCCAUAGAAGUCAUGGUGGAUGCUGUUCACAAAUUAAAAUAUGAAAACUACACCAGCAGCUUCUUCAUCAGAGACAUCAUCAAGCCAGACCCGCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCAGACAAGUGGAAGUUUCCUGGGAGUACCCAGACACGUGGAGCACGCCGCACAGCUACUUCAGCCUCACCUUCUGUGUACAAGUACAAGGCAAGAGCAAGAGAGAGAAGAAAGAUCGUGUCUUCACAGACAAAACCUCGGCGACGGUCAUCUGCAGGAAGAAUGCCUCCAUCUCGGUUCGAGCCCAGGACCGCUACUACAGCAGCAGCUGGAGUGAGUGGGCCUCGGUGCCCUGCAGUGGUGGCGGCGGCGGCGGCAGCAGAAACCUUCCUGUGGCCACGCCGGACCCUGGCAUGUUCCCGUGCCUGCACCACAGCCAAAAUUUACUUCGAGCUGUUUCUAACAUGCUGCAGAAAGCAAGACAAACUUUAGAAUUCUACCCCUGCACCUCAGAAGAAAUAGACCAUGAAGACAUCACCAAAGAUAAAACCAGCACUGUAGAGGCCUGCCUGCCCCUGGAGCUCACCAAGAAUGAAUCCUGCCUCAACAGCAGAGAGACCAGCUUCAUCACCAAUGGCAGCUGCCUGGCCAGCAGGAAAACCAGCUUCAUGAUGGCGCUCUGCCUGAGCAGCAUCUAUGAAGAUUUGAAGAUGUACCAAGUAGAAUUUAAAACCAUGAAUGCCAAGCUGCUCAUGGACCCCAAGAGACAAAUAUUUUUGGAUCAAAACAUGCUGGCUGUCAUUGAUGAGCUCAUGCAAGCAUUAAACUUCAACUCAGAGACGGUGCCCCAGAAGAGCAGCCUGGAGGAGCCAGACUUCUACAAAACCAAGAUCAAGCUCUGCAUCUUAUUACAUGCCUUCCGCAUCCGGGCGGUCACCAUUGACCGUGUCAUGUCCUACUUAAAUGCCAGC 295 hIL12AB_006AUGUGCCACCAGCAGCUGGUGAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCAGCCC ORFCCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGCUGGACUGGUACCCCGACGCCCCCGGCGAGAUGGUGGUGCUGACCUGUGACACCCCCGAGGAGGACGGCAUCACCUGGACCCUGGACCAGAGCAGCGAGGUGCUGGGCAGCGGCAAGACCCUGACCAUCCAGGUGAAGGAGUUCGGGGACGCCGGCCAGUACACCUGCCACAAGGGCGGCGAGGUGCUGAGCCACAGCCUGCUGCUGCUGCACAAGAAGGAGGACGGCAUCUGGAGCACAGAUAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGAGAUGCGAGGCCAAGAACUACAGCGGCAGAUUCACCUGCUGGUGGCUGACCACCAUCAGCACAGACUUGACCUUCAGCGUGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGUGACCUGCGGCGCCGCCACCCUGAGCGCCGAGAGAGUGAGAGGGGACAACAAGGAGUACGAGUACAGCGUGGAGUGCCAGGAGGACAGCGCCUGCCCCGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUGCACAAGCUGAAGUACGAGAACUACACCAGCAGCUUCUUCAUCAGAGACAUCAUCAAGCCCGACCCGCCGAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCAGACAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGAGCACCCCCCACAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGCAAGAGCAAGAGAGAGAAGAAGGACAGAGUGUUCACAGAUAAGACCAGCGCCACCGUGAUCUGCAGAAAGAACGCCAGCAUCAGCGUGAGAGCCCAGGACAGAUACUACAGCAGCAGCUGGAGCGAGUGGGCCAGCGUGCCCUGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCUGCCCGUGGCCACCCCCGACCCCGGCAUGUUCCCCUGCCUGCACCACAGCCAGAACCUGCUGAGAGCCGUGAGCAACAUGCUGCAGAAGGCCAGACAGACCCUGGAGUUCUACCCCUGCACCAGCGAGGAGAUCGACCACGAGGACAUCACCAAGGACAAGACCAGCACCGUGGAGGCCUGCCUGCCCCUGGAGCUGACCAAGAAUGAAAGCUGCCUGAACAGCAGAGAGACCAGCUUCAUCACCAACGGCAGCUGCCUGGCCAGCAGAAAGACCAGCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGAGACAGAUCUUCCUGGACCAGAACAUGCUGGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAGACCGUGCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCAGAAUCAGAGCCGUGACCAUCGACAGAGUGAUGAGCUACCUGAACGCCAGC 296 hIL12AB_007AUGUGCCACCAGCAGCUUGUCAUCUCCUGGUUCUCUCUUGUCUUCCUUGCUUCUCC ORFUCUUGUGGCCAUCUGGGAGCUGAAGAAGGAUGUUUAUGUUGUGGAGUUGGACUGGUACCCUGAUGCUCCUGGAGAAAUGGUGGUUCUCACCUGUGACACUCCUGAGGAGGAUGGCAUCACCUGGACUUUGGACCAGUCUUCUGAGGUUCUUGGCAGUGGAAAAACUCUUACUAUUCAGGUGAAGGAGUUUGGAGAUGCUGGCCAGUACACCUGCCACAAGGGUGGUGAAGUUCUCAGCCACAGUUUACUUCUUCUUCACAAGAAGGAGGAUGGCAUCUGGUCUACUGACAUUUUAAAAGACCAGAAGGAGCCCAAGAACAAGACUUUCCUUCGUUGUGAAGCCAAGAACUACAGUGGUCGUUUCACCUGCUGGUGGCUUACUACUAUUUCUACUGACCUUACUUUCUCUGUGAAGUCUUCUCGUGGCUCUUCUGACCCUCAGGGUGUCACCUGUGGGGCUGCUACUCUUUCUGCUGAGCGUGUGCGUGGGGACAACAAGGAGUAUGAAUACUCGGUGGAGUGCCAGGAGGACUCUGCCUGCCCUGCUGCUGAGGAGUCUCUUCCUAUUGAGGUGAUGGUGGAUGCUGUGCACAAGUUAAAAUAUGAAAACUACACUUCUUCUUUCUUCAUUCGUGACAUUAUAAAACCUGACCCUCCCAAGAACCUUCAGUUAAAACCUUUAAAAAACUCUCGUCAGGUGGAGGUGUCCUGGGAGUACCCUGACACGUGGUCUACUCCUCACUCCUACUUCUCUCUUACUUUCUGUGUCCAGGUGCAGGGCAAGUCCAAGCGUGAGAAGAAGGACCGUGUCUUCACUGACAAGACUUCUGCUACUGUCAUCUGCAGGAAGAAUGCAUCCAUCUCUGUGCGUGCUCAGGACCGUUACUACAGCUCUUCCUGGUCUGAGUGGGCUUCUGUGCCCUGCUCUGGCGGCGGCGGCGGCGGCAGCAGAAAUCUUCCUGUGGCUACUCCUGACCCUGGCAUGUUCCCCUGCCUUCACCACUCGCAGAACCUUCUUCGUGCUGUGAGCAACAUGCUUCAGAAGGCUCGUCAGACUUUAGAAUUCUACCCCUGCACUUCUGAGGAGAUUGACCAUGAAGACAUCACCAAGGACAAGACUUCUACUGUGGAGGCCUGCCUUCCUUUAGAGCUGACCAAGAAUGAAUCCUGCUUAAAUUCUCGUGAGACUUCUUUCAUCACCAAUGGCAGCUGCCUUGCCUCGCGCAAGACUUCUUUCAUGAUGGCUCUUUGCCUUUCUUCCAUCUAUGAAGACUUAAAAAUGUACCAGGUGGAGUUCAAGACCAUGAAUGCAAAGCUUCUCAUGGACCCCAAGCGUCAGAUAUUUUUGGACCAGAACAUGCUUGCUGUCAUUGAUGAGCUCAUGCAGGCUUUAAACUUCAACUCUGAGACUGUGCCUCAGAAGUCUUCUUUAGAAGAGCCUGACUUCUACAAGACCAAGAUAAAACUUUGCAUUCUUCUUCAUGCUUUCCGCAUCCGUGCUGUGACUAUUGACCGUGUGAUGUCCUACUUAAAUGCUUCU 297 hIL12AB_008AUGUGUCAUCAACAACUCGUGAUUAGCUGGUUCAGUCUCGUGUUCCUGGCCUCUCC ORFGCUGGUGGCCAUCUGGGAGCUUAAGAAGGACGUGUACGUGGUGGAGCUCGAUUGGUACCCCGAUGCUCCUGGCGAGAUGGUGGUGCUAACCUGCGAUACCCCCGAGGAGGACGGGAUCACUUGGACCCUGGAUCAGAGUAGCGAAGUCCUGGGCUCUGGCAAGACACUCACAAUCCAGGUGAAGGAAUUCGGAGACGCUGGUCAGUACACUUGCCACAAGGGGGGUGAAGUGCUGUCUCACAGCCUGCUGUUACUGCACAAGAAGGAGGAUGGGAUCUGGUCAACCGACAUCCUGAAGGAUCAGAAGGAGCCUAAGAACAAGACCUUUCUGAGGUGUGAAGCUAAGAACUAUUCCGGAAGAUUCACUUGCUGGUGGUUGACCACAAUCAGCACUGACCUGACCUUUUCCGUGAAGUCCAGCAGAGGAAGCAGCGAUCCUCAGGGCGUAACGUGCGGCGCGGCUACCCUGUCAGCUGAGCGGGUUAGAGGCGACAACAAAGAGUAUGAGUACUCCGUGGAGUGUCAGGAGGACAGCGCCUGCCCCGCAGCCGAGGAGAGUCUGCCCAUCGAGGUGAUGGUGGACGCUGUCCAUAAGUUAAAAUACGAAAAUUACACAAGUUCCUUUUUCAUCCGCGAUAUUAUCAAACCCGAUCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAAUAGCCGACAGGUGGAAGUCUCUUGGGAGUAUCCUGACACCUGGUCCACGCCUCACAGCUACUUUAGUCUGACUUUCUGUGUCCAGGUCCAGGGCAAGAGCAAGAGAGAGAAAAAGGAUAGAGUGUUUACUGACAAGACAUCUGCUACAGUCAUCUGCAGAAAGAACGCCAGUAUCUCAGUGAGGGCGCAGGACAGAUACUACAGUAGUAGCUGGAGCGAAUGGGCUAGCGUGCCCUGUUCAGGGGGCGGCGGAGGGGGCUCCAGGAAUCUGCCCGUGGCCACCCCCGACCCUGGGAUGUUCCCUUGCCUCCAUCACUCACAGAACCUGCUCAGAGCAGUGAGCAACAUGCUCCAAAAGGCCCGCCAGACCCUGGAGUUUUACCCUUGUACUUCAGAAGAGAUCGAUCACGAAGACAUAACAAAGGAUAAAACCAGCACCGUGGAGGCCUGUCUGCCUCUAGAACUCACAAAGAAUGAAAGCUGUCUGAAUUCCAGGGAAACCUCCUUCAUUACUAACGGAAGCUGUCUCGCAUCUCGCAAAACAUCAUUCAUGAUGGCCCUCUGCCUGUCUUCUAUCUAUGAAGAUCUCAAGAUGUAUCAGGUGGAGUUCAAAACAAUGAACGCCAAGCUGCUGAUGGACCCCAAGAGACAGAUCUUCCUGGACCAGAACAUGCUGGCAGUGAUCGAUGAGCUGAUGCAAGCCUUGAACUUCAACUCAGAGACAGUGCCGCAAAAGUCCUCGUUGGAGGAACCAGAUUUUUACAAAACCAAAAUCAAGCUGUGUAUCCUUCUUCACGCCUUUCGGAUCAGAGCCGUGACUAUCGACCGGGUGAUGUCAUACCUGAAUGCUUCC 298 hIL12AB_009AUGUGCCACCAGCAGCUGGUCAUCAGCUGGUUUAGCCUGGUCUUCCUGGCCAGCCC ORFCCUGGUGGCCAUCUGGGAGCUGAAGAAAGAUGUCUAUGUUGUAGAGCUGGACUGGUACCCAGAUGCUCCUGGAGAAAUGGUGGUUCUCACCUGCGACACGCCAGAAGAAGAUGGCAUCACCUGGACGCUGGACCAGAGCAGCGAAGUACUGGGCAGUGGAAAAACGCUGACCAUACAAGUAAAAGAAUUUGGCGAUGCUGGCCAGUACACCUGCCACAAAGGAGGAGAAGUACUGAGCCACAGCCUGCUGCUGCUGCACAAGAAAGAAGAUGGCAUCUGGAGCACCGACAUUUUAAAAGACCAGAAGGAGCCCAAGAACAAAACCUUCCUUCGAUGUGAGGCGAAGAACUACAGUGGCCGCUUCACCUGCUGGUGGCUCACCACCAUCAGCACCGACCUCACCUUCUCGGUGAAGAGCAGCCGUGGUAGCUCAGACCCCCAAGGAGUCACCUGUGGGGCGGCCACGCUGUCGGCAGAAAGAGUUCGAGGCGACAACAAGGAAUAUGAAUACUCGGUGGAAUGUCAAGAAGACUCGGCCUGCCCGGCGGCAGAAGAAAGUCUGCCCAUAGAAGUCAUGGUGGAUGCUGUUCACAAAUUAAAAUAUGAAAACUACACCAGCAGCUUCUUCAUCAGAGACAUCAUCAAGCCAGACCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCAGACAAGUGGAAGUUUCCUGGGAGUACCCAGACACGUGGAGCACGCCGCACAGCUACUUCAGCCUCACCUUCUGUGUACAAGUACAAGGCAAGAGCAAGAGAGAGAAGAAAGAUCGUGUCUUCACCGACAAAACCUCGGCGACGGUCAUCUGCAGGAAGAAUGCAAGCAUCUCGGUUCGAGCCCAGGACCGCUACUACAGCAGCAGCUGGAGUGAGUGGGCCUCGGUGCCCUGCAGUGGUGGCGGCGGCGGCGGCAGCAGAAACCUUCCUGUGGCCACGCCGGACCCUGGCAUGUUUCCGUGCCUGCACCACAGCCAAAAUUUAUUACGAGCUGUUAGCAACAUGCUGCAGAAAGCAAGACAAACUUUAGAAUUCUACCCCUGCACCUCAGAAGAAAUAGACCAUGAAGACAUCACCAAAGAUAAAACCAGCACUGUAGAGGCCUGCCUGCCCCUGGAGCUCACCAAGAACGAGAGCUGCCUCAAUAGCAGAGAGACCAGCUUCAUCACCAAUGGCAGCUGCCUGGCCAGCAGGAAAACCAGCUUCAUGAUGGCGCUCUGCCUGAGCAGCAUCUAUGAAGAUCUGAAGAUGUACCAAGUAGAAUUUAAAACCAUGAAUGCCAAGCUGCUCAUGGACCCCAAGAGACAAAUAUUCCUCGACCAAAACAUGCUGGCUGUCAUUGAUGAGCUCAUGCAAGCAUUAAACUUCAACUCAGAGACGGUGCCCCAGAAGAGCAGCCUGGAGGAGCCAGACUUCUACAAAACCAAGAUCAAGCUCUGCAUCUUAUUACAUGCCUUCCGCAUCCGGGCGGUCACCAUUGACCGUGUCAUGUCCUACUUAAAUGCCAGC 299 hIL12AB_010AUGUGCCACCAGCAGCUUGUCAUCUCCUGGUUUUCUCUUGUCUUCCUCGCUUCUCC ORFUCUUGUGGCCAUCUGGGAGCUGAAGAAAGAUGUCUAUGUUGUAGAGCUGGACUGGUACCCGGACGCUCCUGGAGAAAUGGUGGUUCUCACCUGCGACACUCCUGAAGAAGAUGGCAUCACCUGGACGCUGGACCAAAGCAGCGAAGUUUUAGGCUCUGGAAAAACGCUGACCAUACAAGUAAAAGAAUUUGGCGACGCUGGCCAGUACACGUGCCACAAAGGAGGAGAAGUUUUAAGCCACAGUUUACUUCUUCUUCACAAGAAAGAAGAUGGCAUCUGGAGUACGGACAUUUUAAAAGACCAGAAGGAGCCUAAGAACAAAACCUUCCUCCGCUGUGAAGCUAAGAACUACAGUGGUCGUUUCACCUGCUGGUGGCUCACCACCAUCUCCACUGACCUCACCUUCUCUGUAAAAUCAAGCCGUGGUUCUUCUGACCCCCAAGGAGUCACCUGUGGGGCUGCCACGCUCAGCGCUGAAAGAGUUCGAGGCGACAACAAGGAAUAUGAAUAUUCUGUGGAAUGUCAAGAAGAUUCUGCCUGCCCGGCGGCAGAAGAAAGUCUUCCCAUAGAAGUCAUGGUGGACGCUGUUCACAAAUUAAAAUAUGAAAACUACACCAGCAGCUUCUUCAUUCGUGACAUCAUCAAACCAGACCCUCCUAAGAACCUUCAGUUAAAACCGCUGAAGAACAGCAGACAAGUGGAAGUUUCCUGGGAGUACCCGGACACGUGGAGUACGCCGCACUCCUACUUCAGUUUAACCUUCUGUGUACAAGUACAAGGAAAAUCAAAAAGAGAGAAGAAAGAUCGUGUCUUCACUGACAAAACAUCUGCCACGGUCAUCUGCCGUAAGAACGCUUCCAUCUCGGUUCGAGCCCAGGACCGCUACUACAGCAGCAGCUGGAGUGAGUGGGCAUCUGUUCCCUGCAGUGGUGGCGGCGGCGGCGGCAGCCGCAACCUUCCUGUGGCCACGCCGGACCCUGGCAUGUUCCCGUGCCUUCACCACUCGCAAAAUCUUCUUCGUGCUGUUUCUAACAUGCUGCAGAAGGCGAGACAAACUUUAGAAUUCUACCCGUGCACUUCUGAAGAAAUAGACCAUGAAGACAUCACCAAGGACAAAACCAGCACGGUGGAGGCCUGCCUUCCUUUAGAACUUACUAAGAACGAAAGUUGCCUUAACAGCCGUGAGACCAGCUUCAUCACCAAUGGCAGCUGCCUUGCUAGCAGGAAGACCAGCUUCAUGAUGGCGCUGUGCCUUUCUUCCAUCUAUGAAGAUCUUAAGAUGUACCAAGUAGAAUUUAAAACCAUGAAUGCCAAAUUAUUAAUGGACCCCAAGAGACAAAUAUUCCUCGACCAAAACAUGCUGGCUGUCAUUGAUGAGCUCAUGCAAGCAUUAAACUUCAACUCAGAAACUGUUCCCCAGAAGUCAUCUUUAGAAGAACCGGACUUCUACAAAACAAAAAUAAAACUCUGCAUUCUUCUUCAUGCCUUCCGCAUCCGUGCUGUCACCAUUGACCGUGUCAUGUCCUACUUAAAUGCUUCU 300 hIL12AB_011AUGUGCCACCAGCAGCUGGUGAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCAGCCC ORFCCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGCUGGACUGGUACCCGGACGCGCCGGGGGAGAUGGUGGUGCUGACGUGCGACACGCCGGAGGAGGACGGGAUCACGUGGACGCUGGACCAGAGCAGCGAGGUGCUGGGGAGCGGGAAGACGCUGACGAUCCAGGUGAAGGAGUUCGGGGACGCGGGGCAGUACACGUGCCACAAGGGGGGGGAGGUGCUGAGCCACAGCCUGCUGCUGCUGCACAAGAAGGAGGACGGGAUCUGGAGCACGGACAUCCUGAAGGACCAGAAGGAGCCGAAGAACAAGACGUUCCUGAGGUGCGAGGCGAAGAACUACAGCGGGAGGUUCACGUGCUGGUGGCUGACGACGAUCAGCACGGACCUGACGUUCAGCGUGAAGAGCAGCAGGGGGAGCAGCGACCCGCAGGGGGUGACGUGCGGGGCGGCGACGCUGAGCGCGGAGAGGGUGAGGGGGGACAACAAGGAGUACGAGUACAGCGUGGAGUGCCAGGAGGACAGCGCGUGCCCGGCGGCGGAGGAGAGCCUGCCGAUCGAGGUGAUGGUGGACGCGGUGCACAAGCUGAAGUACGAGAACUACACGAGCAGCUUCUUCAUCAGGGACAUCAUCAAGCCGGACCCGCCGAAGAACCUGCAGCUGAAGCCGCUGAAGAACAGCAGGCAGGUGGAGGUGAGCUGGGAGUACCCGGACACGUGGAGCACGCCGCACAGCUACUUCAGCCUGACGUUCUGCGUGCAGGUGCAGGGGAAGAGCAAGAGGGAGAAGAAGGACAGGGUGUUCACGGACAAGACGAGCGCGACGGUGAUCUGCAGGAAGAACGCGAGCAUCAGCGUGAGGGCGCAGGACAGGUACUACAGCAGCAGCUGGAGCGAGUGGGCGAGCGUGCCGUGCAGCGGGGGGGGGGGGGGGGGGAGCAGGAACCUGCCGGUGGCGACGCCGGACCCGGGGAUGUUCCCGUGCCUGCACCACAGCCAGAACCUGCUGAGGGCGGUGAGCAACAUGCUGCAGAAGGCGAGGCAGACGCUGGAGUUCUACCCGUGCACGAGCGAGGAGAUCGACCACGAGGACAUCACGAAGGACAAGACGAGCACGGUGGAGGCGUGCCUGCCGCUGGAGCUGACGAAGAACGAGAGCUGCCUGAACAGCAGGGAGACGAGCUUCAUCACGAACGGGAGCUGCCUGGCGAGCAGGAAGACGAGCUUCAUGAUGGCGCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACGAUGAACGCGAAGCUGCUGAUGGACCCGAAGAGGCAGAUCUUCCUGGACCAGAACAUGCUGGCGGUGAUCGACGAGCUGAUGCAGGCGCUGAACUUCAACAGCGAGACGGUGCCGCAGAAGAGCAGCCUGGAGGAGCCGGACUUCUACAAGACGAAGAUCAAGCUGUGCAUCCUGCUGCACGCGUUCAGGAUCAGGGCGGUGACGAUCGACAGGGUGAUGAGCUACCUGAACGCGAGC 301 hIL12AB_012AUGUGCCAUCAGCAGCUGGUGAUCAGCUGGUUCAGCCUCGUGUUUCUGGCCAGCCC ORFCCUGGUGGCCAUUUGGGAACUCAAGAAGGACGUGUAUGUAGUGGAACUCGACUGGUACCCUGACGCCCCAGGCGAAAUGGUGGUCUUAACCUGCGACACCCCUGAGGAGGACGGAAUCACCUGGACCUUGGACCAGAGCUCCGAGGUCCUCGGCAGUGGCAAGACCCUGACCAUACAGGUGAAAGAAUUUGGAGACGCAGGGCAAUACACAUGUCACAAGGGCGGGGAGGUUCUUUCUCACUCCCUUCUGCUUCUACAUAAAAAGGAAGACGGAAUUUGGUCUACCGACAUCCUCAAGGACCAAAAGGAGCCUAAGAAUAAAACCUUCUUACGCUGUGAAGCUAAAAACUACAGCGGCAGAUUCACUUGCUGGUGGCUCACCACCAUUUCUACCGACCUGACCUUCUCGGUGAAGUCUUCAAGGGGCUCUAGUGAUCCACAGGGAGUGACAUGCGGGGCCGCCACACUGAGCGCUGAACGGGUGAGGGGCGAUAACAAGGAGUAUGAAUACUCUGUCGAGUGUCAGGAGGAUUCAGCUUGUCCCGCAGCUGAAGAGUCACUCCCCAUAGAGGUUAUGGUCGAUGCUGUGCAUAAACUGAAGUACGAAAACUACACCAGCAGCUUCUUCAUUCGGGACAUUAUAAAACCUGACCCCCCCAAGAACCUGCAACUUAAACCCCUGAAAAACUCUCGGCAGGUCGAAGUUAGCUGGGAGUACCCUGAUACUUGGUCCACCCCCCACUCGUACUUCUCACUGACUUUCUGUGUGCAGGUGCAGGGCAAGAGCAAGAGAGAGAAAAAAGAUCGUGUAUUCACAGACAAGACCUCUGCCACCGUGAUCUGCAGAAAAAACGCUUCCAUCAGUGUCAGAGCCCAAGACCGGUACUAUAGUAGUAGCUGGAGCGAGUGGGCAAGUGUCCCCUGCUCUGGCGGCGGAGGGGGCGGCUCUCGAAACCUCCCCGUCGCUACCCCUGAUCCAGGAAUGUUCCCUUGCCUGCAUCACUCACAGAAUCUGCUGAGAGCGGUCAGCAACAUGCUGCAGAAAGCUAGGCAAACACUGGAGUUUUAUCCUUGUACCUCAGAGGAGAUCGACCACGAGGAUAUUACCAAGGACAAGACCAGCACGGUGGAGGCCUGCUUGCCCCUGGAACUGACAAAGAAUGAAUCCUGCCUUAAUAGCCGUGAGACCUCUUUUAUAACAAACGGAUCCUGCCUGGCCAGCAGGAAGACCUCCUUCAUGAUGGCCCUCUGCCUGUCCUCAAUCUACGAAGACCUGAAGAUGUACCAGGUGGAAUUUAAAACUAUGAACGCCAAGCUGUUGAUGGACCCCAAGCGGCAGAUCUUUCUGGAUCAAAAUAUGCUGGCUGUGAUCGACGAACUGAUGCAGGCCCUCAACUUUAACAGCGAGACCGUGCCACAAAAGAGCAGUCUUGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUCCUUCAUGCCUUCAGGAUAAGAGCUGUCACCAUCGACAGAGUCAUGAGUUACCUGAAUGCAUCC 302 hIL12AB_013AUGUGCCACCAGCAGCUGGUCAUCUCCUGGUUCAGUCUUGUCUUCCUGGCCUCGCC ORFGCUGGUGGCCAUCUGGGAGCUGAAGAAAGAUGUUUAUGUUGUAGAGCUGGACUGGUACCCAGAUGCUCCUGGAGAAAUGGUGGUCCUCACCUGUGACACGCCAGAAGAAGAUGGCAUCACCUGGACGCUGGACCAGAGCAGUGAAGUUCUUGGAAGUGGAAAAACGCUGACCAUACAAGUAAAAGAAUUUGGAGAUGCUGGCCAGUACACCUGCCACAAAGGAGGAGAAGUUCUCAGCCACAGUUUAUUAUUACUUCACAAGAAAGAAGAUGGCAUCUGGUCCACGGACAUUUUAAAAGACCAGAAGGAGCCCAAAAAUAAAACAUUUCUUCGAUGUGAGGCCAAGAACUACAGUGGUCGUUUCACCUGCUGGUGGCUGACCACCAUCUCCACAGACCUCACCUUCAGUGUAAAAAGCAGCCGUGGUUCUUCUGACCCCCAAGGAGUCACCUGUGGGGCUGCCACGCUCUCUGCAGAAAGAGUUCGAGGGGACAACAAAGAAUAUGAGUACUCGGUGGAAUGUCAAGAAGACUCGGCCUGCCCAGCUGCUGAGGAGAGUCUUCCCAUAGAAGUCAUGGUGGAUGCUGUUCACAAAUUAAAAUAUGAAAACUACACCAGCAGCUUCUUCAUCAGAGACAUCAUCAAACCUGACCCGCCCAAGAACUUACAGCUGAAGCCGCUGAAAAACAGCAGACAAGUAGAAGUUUCCUGGGAGUACCCGGACACCUGGUCCACGCCGCACUCCUACUUCUCCCUCACCUUCUGUGUACAAGUACAAGGCAAGAGCAAGAGAGAGAAGAAAGAUCGUGUCUUCACGGACAAAACAUCAGCCACGGUCAUCUGCAGGAAAAAUGCCAGCAUCUCGGUGCGGGCCCAGGACCGCUACUACAGCAGCAGCUGGAGUGAGUGGGCAUCUGUGCCCUGCAGUGGUGGUGGGGGUGGUGGCAGCAGAAACCUUCCUGUGGCCACUCCAGACCCUGGCAUGUUCCCGUGCCUUCACCACUCCCAAAAUUUACUUCGAGCUGUUUCUAACAUGCUGCAGAAAGCAAGACAAACUUUAGAAUUCUACCCGUGCACUUCUGAAGAAAUUGACCAUGAAGACAUCACAAAAGAUAAAACCAGCACAGUGGAGGCCUGUCUUCCUUUAGAGCUGACCAAAAAUGAAUCCUGCCUCAACAGCAGAGAGACCAGCUUCAUCACCAAUGGCAGCUGCCUGGCCUCCAGGAAAACCAGCUUCAUGAUGGCGCUCUGCCUCAGCUCCAUCUAUGAAGAUUUGAAGAUGUACCAAGUAGAAUUUAAAACCAUGAAUGCCAAAUUAUUAAUGGACCCCAAGAGGCAGAUAUUUUUAGAUCAAAACAUGCUGGCAGUUAUUGAUGAGCUCAUGCAAGCAUUAAACUUCAACAGUGAGACUGUACCUCAAAAAAGCAGCCUUGAAGAGCCGGACUUCUACAAAACCAAGAUCAAACUCUGCAUUUUACUUCAUGCCUUCCGCAUCCGGGCGGUCACCAUUGACCGUGUCAUGUCCUACUUAAAUGCCUCG 303 hIL12AB_014AUGUGCCACCAGCAGCUUGUGAUUUCUUGGUUCUCUCUUGUGUUCCUUGCUUCUCC ORFUCUUGUGGCUAUUUGGGAGUUAAAAAAGGACGUGUACGUGGUGGAGCUUGACUGGUACCCUGAUGCUCCUGGCGAGAUGGUGGUGCUUACUUGUGACACUCCUGAGGAGGACGGCAUUACUUGGACUCUUGACCAGUCUUCUGAGGUGCUUGGCUCUGGCAAGACUCUUACUAUUCAGGUGAAGGAGUUCGGGGAUGCUGGCCAGUACACUUGCCACAAGGGCGGCGAGGUGCUUUCUCACUCUCUUCUUCUUCUUCACAAGAAGGAGGACGGCAUUUGGUCUACUGACAUUUUAAAAGACCAGAAGGAGCCCAAGAACAAGACUUUCCUUCGUUGCGAGGCCAAGAACUACUCUGGCCGUUUCACUUGCUGGUGGCUUACUACUAUUUCUACUGACCUUACUUUCUCUGUGAAGUCUUCUCGUGGCUCUUCUGACCCUCAGGGCGUGACUUGUGGGGCUGCUACUCUUUCUGCUGAGCGUGUGCGUGGGGACAACAAGGAGUACGAGUACUCUGUGGAGUGCCAGGAGGACUCUGCUUGCCCUGCUGCUGAGGAGUCUCUUCCUAUUGAGGUGAUGGUGGAUGCUGUGCACAAGUUAAAAUACGAGAACUACACUUCUUCUUUCUUCAUUCGUGACAUUAUUAAGCCUGACCCUCCCAAGAACCUUCAGUUAAAACCUUUAAAAAACUCUCGUCAGGUGGAGGUGUCUUGGGAGUACCCUGACACUUGGUCUACUCCUCACUCUUACUUCUCUCUUACUUUCUGCGUGCAGGUGCAGGGCAAGUCUAAGCGUGAGAAGAAGGACCGUGUGUUCACUGACAAGACUUCUGCUACUGUGAUUUGCAGGAAGAAUGCAUCUAUUUCUGUGCGUGCUCAGGACCGUUACUACUCUUCUUCUUGGUCUGAGUGGGCUUCUGUGCCUUGCUCUGGCGGCGGCGGCGGCGGCUCUAGAAAUCUUCCUGUGGCUACUCCUGACCCUGGCAUGUUCCCUUGCCUUCACCACUCUCAGAACCUUCUUCGUGCUGUGAGCAACAUGCUUCAGAAGGCUCGUCAGACUCUUGAGUUCUACCCUUGCACUUCUGAGGAGAUUGACCACGAGGACAUCACCAAGGACAAGACUUCUACUGUGGAGGCUUGCCUUCCUCUUGAGCUUACCAAGAAUGAAUCUUGCUUAAAUUCUCGUGAGACUUCUUUCAUCACCAACGGCUCUUGCCUUGCCUCGCGCAAGACUUCUUUCAUGAUGGCUCUUUGCCUUUCUUCUAUUUACGAGGACUUAAAAAUGUACCAGGUGGAGUUCAAGACUAUGAAUGCAAAGCUUCUUAUGGACCCCAAGCGUCAGAUUUUCCUUGACCAGAACAUGCUUGCUGUGAUUGACGAGCUUAUGCAGGCUUUAAAUUUCAACUCUGAGACUGUGCCUCAGAAGUCUUCUCUUGAGGAGCCUGACUUCUACAAGACCAAGAUUAAGCUUUGCAUUCUUCUUCAUGCUUUCCGUAUUCGUGCUGUGACUAUUGACCGUGUGAUGUCUUACUUAAAUGCUUCU 304 hIL12AB_015AUGUGUCACCAGCAGCUGGUGAUCAGCUGGUUUAGCCUGGUGUUUCUGGCCAGCCC ORFCCUGGUGGCCAUAUGGGAACUGAAGAAAGAUGUGUAUGUGGUAGAACUGGAUUGGUAUCCGGAUGCCCCCGGCGAAAUGGUGGUGCUGACCUGUGACACCCCCGAAGAAGAUGGUAUCACCUGGACCCUGGACCAGAGCAGCGAGGUGCUGGGCAGCGGCAAAACCCUGACCAUCCAAGUGAAAGAGUUUGGCGAUGCCGGCCAGUACACCUGUCACAAAGGCGGCGAGGUGCUAAGCCAUUCGCUGCUGCUGCUGCACAAAAAGGAAGAUGGCAUCUGGAGCACCGAUAUCCUGAAGGACCAGAAAGAACCCAAAAAUAAGACCUUUCUAAGAUGCGAGGCCAAGAAUUAUAGCGGCCGUUUCACCUGCUGGUGGCUGACGACCAUCAGCACCGAUCUGACCUUCAGCGUGAAAAGCAGCAGAGGCAGCAGCGACCCCCAAGGCGUGACGUGCGGCGCCGCCACCCUGAGCGCCGAGAGAGUGAGAGGCGACAACAAGGAGUAUGAGUACAGCGUGGAGUGCCAGGAGGACAGCGCCUGCCCCGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGAUGCCGUGCACAAGCUGAAGUAUGAAAACUACACCAGCAGCUUCUUCAUCAGAGACAUCAUCAAACCCGACCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAAUAGCAGACAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGAGCACCCCCCAUAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGCAAGAGCAAGAGAGAAAAGAAAGAUAGAGUGUUCACGGACAAGACCAGCGCCACGGUGAUCUGCAGAAAAAAUGCCAGCAUCAGCGUGAGAGCCCAGGACAGAUACUAUAGCAGCAGCUGGAGCGAAUGGGCCAGCGUGCCCUGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCUGCCCGUGGCCACCCCCGACCCCGGCAUGUUCCCCUGCCUGCACCACAGCCAAAACCUGCUGAGAGCCGUGAGCAACAUGCUGCAGAAGGCCAGACAAACCCUGGAAUUUUACCCCUGCACCAGCGAAGAGAUCGAUCAUGAAGAUAUCACCAAAGAUAAAACCAGCACCGUGGAGGCCUGUCUGCCCCUGGAACUGACCAAGAAUGAGAGCUGCCUAAAUAGCAGAGAGACCAGCUUCAUAACCAAUGGCAGCUGCCUGGCCAGCAGAAAGACCAGCUUUAUGAUGGCCCUGUGCCUGAGCAGCAUCUAUGAAGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAAUGCCAAGCUGCUGAUGGAUCCCAAGAGACAGAUCUUUCUGGAUCAAAACAUGCUGGCCGUGAUCGAUGAGCUGAUGCAGGCCCUGAAUUUCAACAGCGAGACCGUGCCCCAAAAAAGCAGCCUGGAAGAACCGGAUUUUUAUAAAACCAAAAUCAAGCUGUGCAUACUGCUGCAUGCCUUCAGAAUCAGAGCCGUGACCAUCGAUAGAGUGAUGAGCUAUCUGAAUGCCAGC 305 hIL12AB_016AUGUGCCACCAGCAGCUGGUCAUCAGCUGGUUCAGCCUGGUCUUCCUGGCCAGCCC ORFCCUGGUGGCCAUCUGGGAGCUGAAGAAGGAUGUUUAUGUUGUGGAGCUGGACUGGUACCCAGAUGCCCCUGGGGAGAUGGUGGUGCUGACCUGUGACACCCCAGAAGAGGAUGGCAUCACCUGGACCCUGGACCAGAGCUCAGAAGUGCUGGGCAGUGGAAAAACCCUGACCAUCCAGGUGAAGGAGUUUGGAGAUGCUGGCCAGUACACCUGCCACAAGGGUGGUGAAGUGCUGAGCCACAGCCUGCUGCUGCUGCACAAGAAGGAGGAUGGCAUCUGGAGCACAGACAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACCUUCCUUCGCUGUGAAGCCAAGAACUACAGUGGCCGCUUCACCUGCUGGUGGCUGACCACCAUCAGCACAGACCUCACCUUCUCGGUGAAGAGCAGCAGAGGCAGCUCAGACCCCCAGGGUGUCACCUGUGGGGCGGCCACGCUGUCGGCGGAGAGAGUUCGAGGGGACAACAAGGAGUAUGAAUACUCGGUGGAGUGCCAGGAGGACUCGGCGUGCCCGGCGGCAGAAGAGAGCCUGCCCAUAGAAGUGAUGGUGGAUGCUGUGCACAAGCUGAAGUAUGAAAACUACACCAGCAGCUUCUUCAUCAGAGACAUCAUCAAGCCAGACCCGCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCAGACAAGUGGAGGUUUCCUGGGAGUACCCAGACACGUGGAGCACCCCCCACAGCUACUUCAGCCUGACCUUCUGUGUCCAGGUGCAGGGCAAGAGCAAGAGAGAGAAGAAGGACAGAGUCUUCACAGACAAGACCUCGGCCACGGUCAUCUGCAGAAAGAAUGCCUCCAUCUCGGUUCGAGCCCAGGACAGAUACUACAGCAGCAGCUGGUCAGAAUGGGCCUCGGUGCCCUGCAGUGGUGGCGGCGGCGGCGGCAGCAGAAACCUGCCUGUUGCCACCCCAGACCCUGGGAUGUUCCCCUGCCUGCACCACAGCCAGAACUUAUUACGAGCUGUUUCUAACAUGCUGCAGAAGGCCAGACAAACCCUGGAGUUCUACCCCUGCACCUCAGAAGAGAUUGACGAUGAAGACAUCACCAAGGACAAGACCAGCACUGUAGAGGCCUGCCUGCCCCUGGAGCUGACCAAGAAUGAAAGCUGCCUGAACAGCAGAGAGACCAGCUUCAUCACCAAUGGAAGCUGCCUGGCCAGCAGAAAGACCAGCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUAUGAAGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAAUGCAAAGCUGCUGAUGGACCCCAAGAGACAAAUAUUUUUGGACCAGAACAUGCUGGCUGUCAUUGAUGAGCUGAUGCAGGCCCUGAACUUCAACUCAGAAACUGUACCCCAGAAGAGCAGCCUGGAGGAGCCAGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUUCAUGCUUUCAGAAUCAGAGCUGUCACCAUUGACCGCGUGAUGAGCUACUUAAAUGCCUCG 306 hIL12AB_017AUGUGCCACCAGCAGCUGGUAAUCAGCUGGUUUUCCCUCGUCUUUCUGGCAUCACC ORFCCUGGUGGCUAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGCUGGAUUGGUACCCUGACGCCCCGGGGGAAAUGGUGGUGUUAACAUGCGACACGCCUGAGGAGGACGGCAUCACCUGGACACUGGACCAGAGCAGCGAGGUGCUUGGGUCUGGUAAAACUCUGACUAUUCAGGUGAAAGAGUUCGGGGAUGCCGGCCAAUAUACUUGCCACAAGGGUGGCGAGGUGCUUUCUCAUUCUCUGCUCCUGCUGCACAAGAAAGAAGAUGGCAUUUGGUCUACUGAUAUUCUGAAAGACCAGAAGGAGCCCAAGAACAAGACCUUUCUGAGAUGCGAGGCUAAAAACUACAGCGGAAGAUUUACCUGCUGGUGGCUGACCACAAUCUCAACCGACCUGACAUUUUCAGUGAAGUCCAGCAGAGGGAGCUCCGACCCUCAGGGCGUGACCUGCGGAGCCGCCACUCUGUCCGCAGAAAGAGUGAGAGGUGAUAAUAAGGAGUACGAGUAUUCAGUCGAGUGCCAAGAGGACUCUGCCUGCCCAGCCGCCGAGGAGAGCCUGCCAAUCGAGGUGAUGGUAGAUGCGGUACACAAGCUGAAGUAUGAGAACUACACAUCCUCCUUCUUCAUAAGAGACAUUAUCAAGCCUGACCCACCUAAAAAUCUGCAACUCAAGCCUUUGAAAAAUUCAAGACAGGUGGAGGUGAGCUGGGAGUACCCUGAUACUUGGAGCACCCCCCAUAGCUACUUUUCGCUGACAUUCUGCGUCCAGGUGCAGGGCAAGUCAAAGAGAGAGAAGAAGGAUCGCGUGUUCACUGAUAAGACAAGCGCCACAGUGAUCUGCAGAAAAAACGCUAGCAUUAGCGUCAGAGCACAGGACCGGUAUUACUCCAGCUCCUGGAGCGAAUGGGCAUCUGUGCCCUGCAGCGGUGGGGGCGGAGGCGGAUCUAGAAACCUCCCCGUUGCCACACCUGAUCCUGGAAUGUUCCCCUGUCUGCACCACAGCCAGAACCUGCUGAGAGCAGUGUCUAACAUGCUCCAGAAGGCCAGGCAGACCCUGGAGUUUUACCCCUGCACCAGCGAGGAAAUCGAUCACGAGGACAUCACCAAAGAUAAAACCUCCACCGUGGAGGCCUGCCUGCCCCUGGAACUGACCAAAAACGAGAGCUGCCUGAAUAGCAGGGAGACCUCCUUCAUCACCAACGGCUCAUGCCUUGCCAGCCGGAAAACUAGCUUCAUGAUGGCCCUGUGCCUGUCUUCGAUCUAUGAGGACCUGAAAAUGUACCAGGUCGAAUUUAAGACGAUGAACGCAAAGCUGCUGAUGGACCCCAAGCGGCAGAUCUUUCUGGACCAGAACAUGCUGGCAGUCAUAGAUGAGUUGAUGCAGGCAUUAAACUUCAACAGCGAGACCGUGCCUCAGAAGUCCAGCCUCGAGGAGCCAGAUUUUUAUAAGACCAAGAUCAAACUAUGCAUCCUGCUGCAUGCUUUCAGGAUUAGAGCCGUCACCAUCGAUCGAGUCAUGUCUUACCUGAAUGCUAGC 307 hIL12AB_018AUGUGUCACCAACAGUUAGUAAUCUCCUGGUUUUCUCUGGUGUUUCUGGCCAGCCC ORFCCUCGUGGCCAUCUGGGAGCUUAAAAAGGAUGUGUACGUGGUGGAGCUGGACUGGUAUCCCGAUGCACCAGGCGAAAUGGUCGUGCUGACCUGCGAUACCCCUGAAGAAGAUGGCAUCACCUGGACUCUGGACCAGUCUUCCGAGGUGCUUGGAUCUGGCAAGACUCUGACAAUACAAGUUAAGGAGUUCGGGGACGCAGGACAGUACACCUGCCACAAAGGCGGCGAGGUCCUGAGUCACUCCCUGUUACUGCUCCACAAGAAAGAGGACGGCAUUUGGUCCACCGACAUUCUGAAGGACCAGAAGGAGCCUAAGAAUAAAACUUUCCUGAGAUGCGAGGCAAAAAACUAUAGCGGCCGCUUUACUUGCUGGUGGCUUACAACAAUCUCUACCGAUUUAACUUUCUCCGUGAAGUCUAGGAGAGGAUCCUCUGAGCCGCAAGGAGUGACUUGCGGAGCCGCCACCUUGAGCGCCGAAAGAGUCCGUGGCGAUAACAAAGAAUACGAGUACUCCGUGGAGUGCCAGGAAGAUUCCGCCUGCCCAGCUGCCGAGGAGUCCCUGCCCAUUGAAGUGAUGGUGGAUGCCGUCCACAAGCUGAAGUACGAAAACUAUACCAGCAGCUUCUUCAUCCGGGAUAUCAUUAAGCCCGACCCUCCUAAAAACCUGCAACUUAAGCCCCUAAAGAAUAGUCGGCAGGUUGAGGUCAGCUGGGAAUAUCCUGACACAUGGAGCACCCCCCACUCUUAUUUCUCCCUGACCUUCUGCGUGCAGGUGCAGGGCAAGAGUAAACGGGAGAAAAAGGACAGGGUCUUUACCGAUAAAACCAGCGCUACGGUUAUCUGUCGGAAGAACGCUUCCAUCUCCGUCCGCGCUCAGGAUCGUUACUACUCGUCCUCAUGGAGCGAGUGGGCCAGCGUGCCCUGCAGCGGCGGCGGUGGAGGCGGAUCCAGAAAUCUGCCUGUUGCCACACCAGACCCUGGCAUGUUCCCCUGUCUGCAUCAUAGCCAGAACCUGCUCAGAGCCGUGAGCAACAUGCUCCAGAAGGCCAGGCAGACAUUGGAGUUCUACCCGUGUACAUCUGAGGAAAUCGAUCACGAAGAUAUAACCAAGGACAAAACCUCUACAGUAGAGGCUUGUUUGCCCCUGGAGUUGACCAAAAACGAGAGUUGCCUGAACAGUCGCGAGACAAGCUUCAUUACUAACGGCAGCUGUCUCGCCUCCAGAAAGACAUCCUUCAUGAUGGCCCUGUGUCUUUCCAGCAUAUACGAAGACCUGAAAAUGUACCAGGUCGAGUUCAAAACAAUGAACGCCAAGCUGCUUAUGGACCCCAAGAGACAGAUCUUCCUCGACCAAAACAUGCUCGCUGUGAUCGAUGAGCUGAUGCAGGCUCUCAACUUCAAUUCCGAAACAGUGCCACAGAAGUCCAGUCUGGAAGAACCCGACUUCUACAAGACCAAGAUUAAGCUGUGUAUUUUGCUGCAUGCGUUUAGAAUCAGAGCCGUGACCAUUGAUCGGGUGAUGAGCUACCUGAACGCCUCG 308 hIL12AB_019AUGUGCCACCAGCAGCUUGUCAUCUCCUGGUUUUCUCUUGUCUUCCUGGCCUCGCC ORFGCUGGUGGCCAUCUGGGAGCUGAAGAAAGAUGUCUAUGUUGUAGAGCUGGACUGGUACCCAGAUGCUCCUGGAGAAAUGGUGGUUCUCACCUGUGACACUCCUGAAGAAGAUGGCAUCACCUGGACGCUGGACCAAAGCUCAGAAGUUCUUGGCAGUGGAAAAACGCUGACCAUACAAGUAAAAGAAUUUGGGGAUGCUGGCCAGUACACGUGCCACAAAGGAGGAGAAGUUCUCAGCCACAGUUUACUUCUUCUUCACAAGAAAGAAGAUGGCAUCUGGUCCACGGACAUUUUAAAAGACCAGAAGGAGCCCAAGAACAAAACCUUCCUCCGCUGUGAGGCCAAGAACUACAGUGGUCGUUUCACCUGCUGGUGGCUCACCACCAUCUCCACUGACCUCACCUUCUCUGUAAAAAGCAGCCGUGGUUCUUCUGACCCCCAAGGAGUCACCUGUGGGGCUGCCACGCUCUCGGCAGAAAGAGUUCGAGGGGACAACAAGGAAUAUGAAUAUUCUGUGGAAUGUCAAGAAGAUUCUGCCUGCCCGGCGGCAGAAGAAAGUCUUCCCAUAGAAGUCAUGGUGGAUGCUGUUCACAAAUUAAAAUAUGAAAACUACACCAGCAGCUUCUUCAUUCGUGACAUCAUCAAACCAGACCCGCCCAAGAACCUUCAGUUAAAACCUUUAAAAAACAGCAGACAAGUAGAAGUUUCCUGGGAGUACCCGGACACGUGGUCCACGCCGCACUCCUACUUCAGUUUAACCUUCUGUGUACAAGUACAAGGAAAAUCAAAAAGAGAGAAGAAAGAUCGUGUCUUCACUGACAAAACAUCUGCCACGGUCAUCUGCAGGAAGAAUGCCUCCAUCUCGGUUCGAGCCCAGGACCGCUACUACAGCAGCAGCUGGAGUGAGUGGGCAUCUGUUCCCUGCAGUGGUGGCGGCGGCGGCGGCAGCCGCAACCUUCCUGUGGCCACGCCGGACCCUGGCAUGUUCCCGUGCCUUCACCACUCCCAAAAUCUUCUUCGUGCUGUUUCUAACAUGCUGCAGAAGGCGCGCCAAACUUUAGAAUUCUACCCGUGCACUUCUGAAGAAAUAGACCAUGAAGACAUCACCAAAGAUAAAACCAGCACGGUGGAGGCCUGCCUUCCUUUAGAGCUGACCAAGAAUGAAUCCUGCCUCAACAGCAGAGAGACCAGCUUCAUCACCAAUGGCAGCUGCCUGGCCUCGCGCAAGACCAGCUUCAUGAUGGCGCUGUGCCUUUCUUCCAUCUAUGAAGAUUUAAAGAUGUACCAAGUAGAAUUUAAAACCAUGAAUGCCAAAUUAUUAAUGGACCCCAAAAGACAAAUAUUUUUGGAUCAAAACAUGCUGGCUGUCAUUGAUGAGCUCAUGCAAGCAUUAAACUUCAACUCAGAAACUGUUCCCCAGAAGUCAUCUUUAGAAGAGCCGGACUUCUACAAAACAAAAAUAAAACUCUGCAUUCUUCUUCAUGCCUUCCGCAUCCGUGCUGUCACCAUUGACCGUGUCAUGUCCUACUUAAAUGCUUCU 309 hIL12AB_020AUGUGCCACCAGCAGCUGGUGAUCAGCUGGUUCAGCCUGGUGUUCCUGGCUAGCCC ORFUCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGUUAGACUGGUACCCCGACGCUCCCGGCGAGAUGGUGGUGCUGACCUGCGACACCCCCGAGGAGGACGGGAUCACCUGGACCCUGGAUCAGUCAAGCGAGGUGCUGGGAAGCGGCAAGACCCUGACCAUCCAGGUGAAGGAGUUCGGCGACGCCGGCCAAUACACUUGCCACAAGGGAGGCGAGGUGCUGUCCCACUCCCUCCUGCUGCUGCACAAAAAGGAAGACGGCAUCUGGAGCACCGACAUCCUGAAAGACCAGAAGGAGCCUAAGAACAAGACAUUCCUCAGAUGCGAGGCCAAGAAUUACUCCGGGAGAUUCACCUGUUGGUGGCUGACCACCAUCAGCACAGACCUGACCUUCAGCGUGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGUGACCUGUGGCGCCGCCACCCUGAGCGCCGAAAGAGUGCGCGGCGACAACAAGGAGUACGAGUACUCCGUGGAAUGCCAGGAGGACAGCGCCUGCCCCGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUCCACAAGCUGAAGUACGAGAACUACACCUCUAGCUUCUUCAUCCGGGACAUCAUCAAGCCCGAUCCCCCCAAGAACCUGCAGCUGAAACCCCUGAAGAACAGCAGACAGGUGGAGGUGAGCUGGGAGUAUCCCGACACCUGGUCCACCCCCCACAGCUAUUUUAGCCUGACCUUCUGCGUGCAAGUGCAGGGCAAGAGCAAGAGAGAGAAGAAGGACCGCGUGUUCACCGACAAAACCAGCGCCACCGUGAUCUGCAGAAAGAACGCCAGCAUCAGCGUGAGGGCCCAGGAUAGAUACUACAGUUCCAGCUGGAGCGAGUGGGCCAGCGUGCCCUGCAGCGGCGGCGGCGGGGGAGGCUCUAGAAACCUGCCCGUGGCUACCCCCGAUCCCGGAAUGUUCCCCUGCCUGCACCACAGCCAGAACCUGCUGAGGGCGGUGUCCAACAUGCUUCAGAAGGCCCGGCAGACCCUGGAGUUCUACCCCUGUACCUCUGAGGAGAUCGAUCAUGAGGACAUCACAAAGGACAAAACCAGCACCGUGGAGGCCUGCCUGCCCCUGGAGCUGACCAAGAACGAGAGCUGCCUGAACUCCCGCGAGACCAGCUUCAUCACGAACGGCAGCUGCCUGGCCAGCAGGAAGACCUCCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAAAUGUACCAGGUGGAGUUUAAGACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGGCAAAUCUUCCUGGACCAGAACAUGCUGGCAGUGAUCGACGAGCUCAUGCAGGCCCUGAACUUCAAUAGCGAGACAGUCCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUUUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUUAGAAUCCGUGCCGUGACCAUUGACAGAGUGAUGAGCUACCUGAAUGCCAGC 310 hIL12AB_021AUGUGCCACCAGCAGCUGGUGAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCAGCCC ORFUCUGGUUGCCAUCUGGGAGCUGAAGAAAGACGUGUACGUCGUGGAACUGGACUGGUAUCCGGACGCCCCGGGCGAGAUGGUGGUGCUGACCUGUGACACCCCCGAGGAGGACGGCAUCACCUGGACGCUGGACCAAUCCUCCGAGGUGCUGGGAAGCGGCAAGACCCUGACCAUCCAGGUGAAGGAAUUCGGGGACGCCGGGCAGUACACCUGCCACAAGGGGGGCGAAGUGCUGUCCCACUCGCUGCUGCUCCUGCAUAAGAAGGAGGAUGGAAUCUGGUCCACCGACAUCCUCAAAGAUCAGAAGGAGCCCAAGAACAAGACGUUCCUGCGCUGUGAAGCCAAGAAUUAUUCGGGGCGAUUCACGUGCUGGUGGCUGACAACCAUCAGCACCGACCUGACGUUUAGCGUGAAGAGCAGCAGGGGGUCCAGCGACCCCCAGGGCGUGACGUGCGGCGCCGCCACCCUCUCCGCCGAGAGGGUGCGGGGGGACAAUAAGGAGUACGAGUACAGCGUGGAAUGCCAGGAGGACAGCGCCUGCCCCGCCGCGGAGGAAAGCCUCCCGAUAGAGGUGAUGGUGGACGCCGUGCACAAGCUCAAGUAUGAGAAUUACACCAGCAGCUUUUUCAUCCGGGACAUUAUCAAGCCCGACCCCCCGAAGAACCUCCAGCUGAAGCCCCUGAAGAACAGCCGGCAGGUGGAAGUCUCCUGGGAGUAUCCCGACACCUGGAGCACCCCGCACAGCUACUUCUCCCUGACCUUCUGUGUGCAGGUGCAGGGCAAGUCCAAGAGGGAAAAGAAGGACAGGGUUUUCACCGACAAGACCAGCGCGACCGUGAUCUGCCGGAAGAACGCCAGCAUAAGCGUCCGCGCCCAAGAUAGGUACUACAGCAGCUCCUGGAGCGAGUGGGCUAGCGUGCCCUGCAGCGGGGGCGGGGGUGGGGGCUCCAGGAACCUGCCAGUGGCGACCCCCGACCCCGGCAUGUUCCCCUGCCUCCAUCACAGCCAGAACCUGCUGAGGGCCGUCAGCAAUAUGCUGCAGAAGGCCAGGCAGACCCUGGAAUUCUACCCCUGCACGUCGGAGGAGAUCGAUCACGAGGAUAUCACAAAAGACAAGACUUCCACCGUGGAGGCCUGCCUGCCCCUGGAGCUCACCAAGAAUGAGUCCUGUCUGAACUCCCGGGAAACCAGCUUCAUCACCAACGGGUCCUGCCUGGCCAGCAGGAAGACCAGCUUUAUGAUGGCCCUGUGCCUGUCGAGCAUCUACGAGGACCUGAAGAUGUACCAGGUCGAGUUCAAGACAAUGAACGCCAAGCUGCUGAUGGACCCCAAGAGGCAAAUCUUCCUGGACCAGAAUAUGCUUGCCGUCAUCGACGAGCUCAUGCAGGCCCUGAACUUCAACUCCGAGACCGUGCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCGUUCAGGAUCCGGGCAGUCACCAUCGACCGUGUGAUGUCCUACCUGAACGCCAGC 311 hIL12AB_022AUGUGCCAUCAGCAGCUGGUGAUCAGCUGGUUCAGCCUGGUGUUCCUCGCCUCUCC ORFCCUGGUGGCCAUCUGGGAGCUCAAAAAGGACGUGUACGUGGUGGAGCUCGACUGGUACCCAGACGCCCCCGGGGAGAUGGUGGUGCUGACCUGCGACACCCCCGAAGAAGACGGCAUCACGUGGACCCUCGACCAGUCCAGCGAGGUGCUGGGGAGCGGGAAGACUCUGACCAUCCAGGUCAAGGAGUUCGGGGACGCCGGGCAGUACACGUGCCACAAGGGCGGCGAAGUCUUAAGCCACAGCCUGCUCCUGCUGCACAAGAAGGAGGACGGGAUCUGGUCCACAGACAUACUGAAGGACCAGAAGGAGCCGAAGAAUAAAACCUUUCUGAGGUGCGAGGCCAAGAACUAUUCCGGCAGGUUCACGUGCUGGUGGCUUACAACAAUCAGCACAGACCUGACGUUCAGCGUGAAGUCCAGCCGCGGCAGCAGCGACCCCCAGGGGGUGACCUGCGGCGCCGCCACCCUGAGCGCCGAGCGGGUGCGCGGGGACAACAAGGAGUACGAGUACUCCGUGGAGUGCCAGGAAGACAGCGCCUGUCCCGCCGCCGAAGAGAGCCUGCCUAUCGAGGUCAUGGUAGAUGCAGUGCAUAAGCUGAAGUACGAGAACUAUACGAGCAGCUUUUUCAUACGCGACAUCAUCAAGCCCGACCCCCCCAAGAACCUGCAGCUUAAGCCCCUGAAGAAUAGCCGGCAGGUGGAGGUCUCCUGGGAGUACCCCGACACCUGGUCAACGCCCCACAGCUACUUCUCCCUGACCUUUUGUGUCCAAGUCCAGGGAAAGAGCAAGAGGGAGAAGAAAGAUCGGGUGUUCACCGACAAGACCUCCGCCACGGUGAUCUGCAGGAAGAACGCCAGCAUCUCCGUGAGGGCGCAAGACAGGUACUACUCCAGCAGCUGGUCCGAAUGGGCCAGCGUGCCCUGCUCCGGCGGCGGGGGCGGCGGCAGCCGAAACCUACCCGUGGCCACGCCGGAUCCCGGCAUGUUUCCCUGCCUGCACCACAGCCAGAACCUCCUGAGGGCCGUGUCCAACAUGCUGCAGAAGGCCAGGCAGACUCUGGAGUUCUACCCCUGCACGAGCGAGGAGAUCGAUCACGAGGACAUCACCAAGGAUAAGACCAGCACUGUGGAGGCCUGCCUUCCCCUGGAGCUGACCAAGAACGAGAGCUGUCUGAACUCCAGGGAGACCUCAUUCAUCACCAACGGCUCCUGCCUGGCCAGCAGGAAAACCAGCUUCAUGAUGGCCUUGUGUCUCAGCUCCAUCUACGAGGACCUGAAGAUGUAUCAGGUCGAGUUCAAGACAAUGAACGCCAAGCUGCUGAUGGACCCCAAAAGGCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUCAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAGACGGUGCCCCAGAAAAGCUCCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCAGGAUCAGGGCAGUGACCAUCGACCGGGUGAUGUCAUACCUUAACGCCAGC 312 hIL12AB_023AUGUGCCAUCAGCAGCUGGUGAUCUCCUGGUUCAGCCUGGUGUUUCUGGCCUCGCC ORFCCUGGUCGCCAUCUGGGAGCUGAAGAAAGACGUGUACGUCGUCGAACUGGACUGGUACCCCGACGCCCCCGGGGAGAUGGUGGUGCUGACCUGCGACACGCCGGAGGAGGACGGCAUCACCUGGACCCUGGAUCAAAGCAGCGAGGUGCUGGGCAGCGGCAAGACCCUGACCAUCCAAGUGAAGGAAUUCGGCGAUGCCGGCCAGUACACCUGUCACAAAGGGGGCGAGGUGCUCAGCCACAGCCUGCUGCUGCUGCACAAGAAGGAGGAUGGCAUCUGGAGCACCGAUAUCCUGAAGGACCAGAAAGAGCCCAAGAACAAGACGUUCCUGAGGUGCGAGGCCAAGAACUACAGCGGUAGGUUCACGUGUUGGUGGCUGACCACCAUCAGCACCGACCUGACGUUCAGCGUGAAGAGCUCCAGGGGCAGCUCCGACCCACAGGGGGUGACGUGCGGGGCCGCAACCCUCAGCGCCGAAAGGGUGCGGGGGGACAACAAGGAGUACGAAUACUCCGUGGAGUGCCAGGAAGAUUCGGCCUGCCCCGCCGCGGAGGAGAGCCUCCCCAUCGAGGUAAUGGUGGACGCCGUGCAUAAGCUGAAGUACGAGAACUACACCAGCUCGUUCUUCAUCCGAGACAUCAUCAAACCCGACCCGCCCAAAAAUCUGCAGCUCAAGCCCCUGAAGAACUCCAGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGUCCACCCCGCACAGCUACUUCUCCCUGACAUUCUGCGUGCAGGUGCAGGGCAAGAGCAAGCGGGAGAAGAAGGACAGGGUGUUCACCGACAAGACGAGCGCCACCGUGAUCUGCCGAAAGAACGCCAGCAUCUCGGUGCGCGCCCAGGAUAGGUACUAUUCCAGCUCCUGGAGCGAGUGGGCCUCGGUACCCUGCAGCGGCGGCGGGGGCGGCGGCAGUAGGAAUCUGCCCGUGGCUACCCCGGACCCGGGCAUGUUCCCCUGCCUCCACCACAGCCAGAACCUGCUGAGGGCCGUGAGCAACAUGCUGCAGAAGGCCAGACAGACGCUGGAGUUCUACCCCUGCACGAGCGAGGAGAUCGACCACGAGGACAUCACCAAGGAUAAAACUUCCACCGUCGAGGCCUGCCUGCCCUUGGAGCUGACCAAGAAUGAAUCCUGUCUGAACAGCAGGGAGACCUCGUUUAUCACCAAUGGCAGCUGCCUCGCCUCCAGGAAGACCAGCUUCAUGAUGGCCCUCUGUCUGAGCUCCAUCUAUGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCGAAGCUGCUGAUGGACCCCAAGAGGCAGAUCUUCCUGGAUCAGAAUAUGCUGGCGGUGAUCGACGAGCUCAUGCAGGCCCUCAAUUUCAAUAGCGAGACAGUGCCCCAGAAGUCCUCCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGUAUCCUGCUGCACGCCUUCCGGAUCCGGGCCGUCACCAUCGACCGGGUCAUGAGCUACCUCAAUGCCAGC 313 hIL12AB_024AUGUGCCACCAGCAGCUGGUGAUCUCCUGGUUCUCCCUGGUGUUCCUGGCCUCGCC ORFCCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUCGUGGAGCUCGACUGGUACCCCGACGCCCCUGGCGAGAUGGUGGUGCUGACCUGCGACACCCCAGAGGAGGAUGGCAUCACCUGGACCCUGGAUCAGUCCUCCGAGGUGCUGGGCUCCGGCAAGACGCUGACCAUCCAAGUGAAGGAGUUCGGUGACGCCGGACAGUAUACCUGCCAUAAGGGCGGCGAGGUCCUGUCCCACAGCCUCCUCCUCCUGCAUAAGAAGGAGGACGGCAUCUGGAGCACCGACAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACCUUUCUGAGGUGCGAGGCCAAGAACUACAGCGGCCGAUUCACCUGCUGGUGGCUCACCACCAUAUCCACCGACCUGACUUUCUCCGUCAAGUCCUCCCGGGGGUCCAGCGACCCCCAGGGAGUGACCUGCGGCGCCGCCACCCUCAGCGCCGAGCGGGUGCGGGGGGACAACAAGGAGUACGAAUACUCCGUCGAGUGCCAGGAGGACUCCGCCUGCCCGGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUCGACGCGGUGCACAAGCUGAAGUACGAGAACUACACCAGCAGUUUCUUCAUCAGGGAUAUCAUCAAGCCAGAUCCCCCGAAGAAUCUGCAACUGAAGCCGCUGAAAAACUCACGACAGGUGGAGGUGAGCUGGGAGUACCCCGACACGUGGAGCACCCCACAUUCCUACUUCAGCCUGACCUUCUGCGUGCAGGUCCAGGGCAAGAGCAAGCGGGAGAAGAAGGACAGGGUGUUCACGGAUAAGACCAGUGCCACCGUGAUCUGCAGGAAGAACGCCUCUAUUAGCGUGAGGGCCCAGGAUCGGUAUUACUCCUCGAGCUGGAGCGAAUGGGCCUCCGUGCCCUGCAGUGGGGGGGGUGGAGGCGGGAGCAGGAACCUGCCCGUAGCAACCCCCGACCCCGGGAUGUUCCCCUGUCUGCACCACUCGCAGAACCUGCUGCGCGCGGUGAGCAACAUGCUCCAAAAAGCCCGUCAGACCUUAGAGUUCUACCCCUGCACCAGCGAAGAAAUCGACCACGAAGACAUCACCAAGGACAAAACCAGCACCGUGGAGGCGUGCCUGCCGCUGGAGCUGACCAAGAACGAGAGCUGCCUCAACUCCAGGGAGACCAGCUUUAUCACCAACGGCUCGUGCCUAGCCAGCCGGAAAACCAGCUUCAUGAUGGCCCUGUGCCUGAGCUCCAUUUACGAGGACCUGAAGAUGUAUCAGGUGGAGUUCAAGACCAUGAAUGCCAAACUCCUGAUGGACCCCAAGAGGCAGAUCUUCCUGGACCAGAACAUGCUCGCGGUGAUCGAUGAGCUGAUGCAGGCCCUGAACUUUAAUAGCGAGACCGUGCCCCAGAAAAGCAGCCUGGAGGAGCCGGACUUCUACAAGACCAAAAUCAAGCUGUGCAUCCUGCUCCACGCCUUCCGCAUCCGGGCCGUGACCAUCGACAGGGUGAUGAGCUACCUGAACGCCAGC 314 hIL12AB_025AUGUGCCAUCAGCAGCUGGUGAUUUCCUGGUUCUCCCUGGUGUUCCUGGCCAGCCC ORFCCUCGUGGCGAUCUGGGAGCUAAAGAAGGACGUGUACGUGGUGGAGCUGGACUGGUACCCGGACGCACCCGGCGAGAUGGUCGUUCUGACCUGCGAUACGCCAGAGGAGGACGGCAUCACCUGGACCCUCGAUCAGAGCAGCGAGGUCCUGGGGAGCGGAAAGACCCUGACCAUCCAGGUCAAGGAGUUCGGCGACGCCGGCCAGUACACCUGCCACAAAGGUGGCGAGGUCCUGAGCCACUCGCUGCUGCUCCUGCAUAAGAAGGAGGACGGAAUCUGGAGCACAGACAUCCUGAAAGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGAGGUGCGAGGCCAAGAACUACAGCGGGCGCUUCACGUGCUGGUGGCUGACCACCAUCAGCACGGACCUCACCUUCUCCGUGAAGAGCAGCCGGGGAUCCAGCGAUCCCCAAGGCGUCACCUGCGGCGCGGCCACCCUGAGCGCGGAGAGGGUCAGGGGCGAUAAUAAGGAGUAUGAGUACAGCGUGGAGUGCCAGGAGGACAGCGCCUGCCCGGCCGCCGAGGAGUCCCUGCCAAUCGAAGUGAUGGUCGACGCCGUGCACAAGCUGAAGUACGAGAACUACACCAGCAGCUUCUUCAUCCGGGAUAUCAUCAAGCCCGAUCCCCCGAAGAACCUGCAGCUGAAGCCCCUCAAGAACAGCCGGCAGGUGGAGGUGAGUUGGGAGUACCCCGACACCUGGUCAACGCCCCACAGCUACUUCUCCCUGACCUUCUGUGUGCAGGUGCAGGGAAAGAGCAAGAGGGAGAAGAAAGACCGGGUCUUCACCGACAAGACCAGCGCCACGGUGAUCUGCAGGAAGAACGCAAGCAUCUCCGUGAGGGCCCAGGACAGGUACUACAGCUCCAGCUGGUCCGAAUGGGCCAGCGUGCCCUGUAGCGGCGGCGGGGGCGGUGGCAGCCGCAACCUCCCAGUGGCCACCCCCGACCCCGGCAUGUUCCCCUGCCUGCACCACAGCCAGAAUCUGCUGAGGGCCGUGAGUAACAUGCUGCAGAAGGCAAGGCAAACCCUCGAAUUCUAUCCCUGCACCUCCGAGGAGAUCGACCACGAGGAUAUCACCAAGGACAAGACCAGCACCGUCGAGGCCUGUCUCCCCCUGGAGCUGACCAAGAAUGAGAGCUGCCUGAACAGCCGGGAGACCAGCUUCAUCACCAACGGGAGCUGCCUGGCCUCCAGGAAGACCUCGUUCAUGAUGGCGCUGUGCCUCUCAAGCAUAUACGAGGAUCUGAAGAUGUACCAGGUGGAGUUUAAGACGAUGAACGCCAAGCUGCUGAUGGACCCGAAGAGGCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUGAUAGACGAGCUCAUGCAGGCCCUGAACUUCAACUCCGAGACCGUGCCGCAGAAGUCAUCCCUCGAGGAGCCCGACUUCUAUAAGACCAAGAUCAAGCUGUGCAUCCUGCUCCACGCCUUCCGGAUAAGGGCCGUGACGAUCGACAGGGUGAUGAGCUACCUUAACGCCAGC 315 hIL12AB_026AUGUGCCACCAGCAGCUCGUGAUCAGCUGGUUCUCCCUGGUGUUUCUCGCCAGCCC ORFCCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGCUGGACUGGUACCCUGACGCCCCGGGGGAGAUGGUCGUGCUGACCUGCGACACCCCCGAAGAGGACGGUAUCACCUGGACCCUGGACCAGUCCAGCGAGGUGCUGGGCAGCGGCAAGACCCUGACUAUUCAAGUCAAGGAGUUCGGAGACGCCGGCCAGUACACCUGCCACAAGGGUGGAGAGGUGUUAUCACACAGCCUGCUGCUGCUGCACAAGAAGGAAGACGGGAUCUGGAGCACCGACAUCCUGAAGGACCAGAAGGAGCCCAAAAACAAGACCUUCCUGCGGUGCGAGGCCAAGAACUAUUCGGGCCGCUUUACGUGCUGGUGGCUGACCACCAUCAGCACUGAUCUCACCUUCAGCGUGAAGUCCUCCCGGGGGUCGUCCGACCCCCAGGGGGUGACCUGCGGGGCCGCCACCCUGUCCGCCGAGAGAGUGAGGGGCGAUAAUAAGGAGUACGAGUACAGCGUUGAGUGCCAGGAAGAUAGCGCCUGUCCCGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUCCACAAGCUGAAGUAUGAGAACUACACCUCAAGCUUCUUCAUCAGGGACAUCAUCAAACCCGAUCCGCCCAAGAAUCUGCAGCUGAAGCCCCUGAAAAAUAGCAGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGUCCACCCCCCAUAGCUAUUUCUCCCUGACGUUCUGCGUGCAGGUGCAAGGGAAGAGCAAGCGGGAGAAGAAGGACCGGGUGUUCACCGACAAGACCUCCGCCACCGUGAUCUGUAGGAAGAACGCGUCGAUCUCGGUCAGGGCCCAGGACAGGUAUUACAGCAGCAGCUGGAGCGAGUGGGCGAGCGUGCCCUGCUCGGGCGGCGGCGGCGGCGGGAGCAGAAAUCUGCCCGUGGCCACCCCAGACCCCGGAAUGUUCCCCUGCCUGCACCAUUCGCAGAACCUCCUGAGGGCCGUGAGCAACAUGCUGCAGAAGGCCCGCCAGACGCUGGAGUUCUACCCCUGCACGAGCGAGGAGAUCGACCACGAAGACAUCACCAAGGACAAAACCAGCACCGUGGAGGCCUGCCUGCCCCUGGAGCUGACCAAAAACGAAUCCUGCCUCAACAGCCGGGAGACCAGCUUCAUCACCAACGGCAGCUGCCUGGCCAGCCGAAAGACCUCCUUCAUGAUGGCCCUCUGCCUGAGCAGCAUCUAUGAGGAUCUGAAGAUGUAUCAGGUGGAGUUCAAGACCAUGAAUGCCAAGCUGCUGAUGGACCCCAAGAGGCAGAUAUUCCUGGACCAGAAUAUGCUGGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAGACCGUCCCCCAGAAGUCCAGCCUGGAGGAGCCGGACUUUUACAAAACGAAGAUCAAGCUGUGCAUACUGCUGCACGCCUUCAGGAUCCGGGCCGUGACAAUCGACAGGGUGAUGUCCUACCUGAACGCCAGC 316 hIL12AB_027AUGUGUCACCAGCAGCUGGUGAUCAGCUGGUUCUCCCUGGUGUUCCUGGCCAGCCC ORFCCUGGUGGCCAUCUGGGAGCUCAAGAAGGACGUCUACGUCGUGGAGCUGGAUUGGUACCCCGACGCUCCCGGGGAGAUGGUGGUGCUGACCUGCGACACCCCCGAGGAGGACGGCAUCACCUGGACGCUGGACCAGAGCUCAGAGGUGCUGGGAAGCGGAAAGACACUGACCAUCCAGGUGAAGGAGUUCGGGGAUGCCGGGCAGUAUACCUGCCACAAGGGCGGCGAAGUGCUGAGCCAUUCCCUGCUGCUGCUGCACAAGAAGGAGGACGGCAUAUGGUCCACCGACAUCCUGAAGGAUCAGAAGGAGCCGAAGAAUAAAACCUUCCUGAGGUGCGAGGCCAAGAAUUACAGCGGCCGAUUCACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUGACCUUCAGUGUGAAGUCCUCACGGGGCAGCUCAGAUCCCCAGGGCGUGACCUGCGGGGCCGCGACACUCAGCGCCGAGCGGGUGAGGGGUGAUAACAAGGAGUACGAGUAUUCUGUGGAGUGCCAGGAAGACUCCGCCUGUCCCGCCGCCGAGGAGUCCCUGCCCAUCGAGGUGAUGGUGGACGCCGUGCAUAAACUGAAGUACGAGAACUACACCUCCAGCUUCUUCAUCCGGGAUAUAAUCAAGCCCGACCCUCCGAAAAACCUGCAGCUGAAGCCCCUUAAAAACAGCCGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGAGCACCCCCCAUAGCUAUUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGGAAGUCCAAGCGCGAGAAAAAGGACCGGGUGUUCACCGACAAGACGAGCGCCACCGUGAUCUGCCGGAAGAACGCCAGUAUAAGCGUAAGGGCCCAGGAUAGGUACUACAGCUCCAGCUGGUCGGAGUGGGCCUCCGUGCCCUGUUCCGGCGGCGGGGGGGGUGGCAGCAGGAACCUCCCCGUGGCCACGCCGGACCCCGGCAUGUUCCCGUGCCUGCACCACUCCCAAAACCUCCUGCGGGCCGUCAGCAACAUGCUGCAAAAGGCGCGGCAGACCCUGGAGUUUUACCCCUGUACCUCCGAAGAGAUCGACCACGAGGAUAUCACCAAGGAUAAGACCUCCACCGUGGAGGCCUGUCUCCCCCUGGAGCUGACCAAGAACGAGAGCUGUCUUAACAGCAGAGAGACCUCGUUCAUAACGAACGGCUCCUGCCUCGCUUCCAGGAAGACGUCGUUCAUGAUGGCGCUGUGCCUGUCCAGCAUCUACGAGGACCUGAAGAUGUAUCAGGUCGAGUUCAAAACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGAGGCAGAUCUUCCUGGACCAGAACAUGCUCGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAAACCGUGCCCCAGAAGUCAAGCCUGGAGGAGCCGGACUUCUAUAAGACCAAGAUCAAGCUGUGUAUCCUGCUACACGCUUUUCGUAUCCGGGCCGUGACCAUCGACAGGGUUAUGUCGUACUUGAACGCCAGC 317 hIL12AB_028AUGUGCCACCAACAGCUCGUGAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCAGCCC ORFGCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGCUGGACUGGUACCCCGACGCCCCCGGCGAGAUGGUGGUCCUGACCUGCGACACGCCGGAAGAGGACGGCAUCACCUGGACCCUGGAUCAGUCCAGCGAGGUGCUGGGCUCCGGCAAGACCCUGACCAUUCAGGUGAAGGAGUUCGGCGACGCCGGUCAGUACACCUGCCACAAGGGCGGCGAGGUGCUGAGCCACAGCCUACUGCUCCUGCACAAAAAGGAGGAUGGAAUCUGGUCCACCGACAUCCUCAAGGACCAGAAGGAGCCGAAGAACAAGACGUUCCUCCGGUGCGAGGCCAAGAACUACAGCGGCAGGUUUACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUGACAUUUUCCGUGAAGAGCAGCCGCGGCAGCAGCGAUCCCCAGGGCGUGACCUGCGGGGCGGCCACCCUGUCCGCCGAGCGUGUGAGGGGCGACAACAAGGAGUACGAGUACAGCGUGGAAUGCCAGGAGGACAGCGCCUGUCCCGCCGCCGAGGAGAGCCUGCCAAUCGAGGUCAUGGUGGACGCCGUGCACAAGCUGAAGUACGAGAACUACACGAGCAGCUUCUUCAUCAGGGACAUCAUCAAACCGGACCCGCCCAAGAACCUGCAGCUGAAACCCUUGAAAAACAGCAGGCAGGUGGAAGUGUCUUGGGAGUACCCCGACACCUGGUCCACCCCCCACAGCUACUUUAGCCUGACCUUCUGUGUGCAGGUCCAGGGCAAGUCCAAGAGGGAGAAGAAGGACAGGGUGUUCACCGACAAAACCAGCGCCACCGUGAUCUGCAGGAAGAACGCCUCCAUCAGCGUGCGGGCCCAGGACAGGUAUUACAGCUCGUCGUGGAGCGAGUGGGCCAGCGUGCCCUGCUCCGGGGGAGGCGGCGGCGGAAGCCGGAAUCUGCCCGUGGCCACCCCCGAUCCCGGCAUGUUCCCGUGUCUGCACCACAGCCAGAACCUGCUGCGGGCCGUGAGCAACAUGCUGCAGAAGGCCCGCCAAACCCUGGAGUUCUACCCCUGUACAAGCGAGGAGAUCGACCAUGAGGACAUUACCAAGGACAAGACCAGCACCGUGGAGGCCUGCCUGCCCCUCGAGCUCACAAAGAACGAAUCCUGCCUGAAUAGCCGCGAGACCAGCUUUAUCACGAACGGGUCCUGCCUCGCCAGCCGGAAGACAAGCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAAAUGUACCAAGUGGAGUUCAAAACGAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGCCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUCAUCGACGAGCUCAUGCAGGCCCUGAACUUCAACAGCGAGACCGUGCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUCUACAAGACGAAGAUCAAGCUCUGCAUCCUGCUGCACGCUUUCCGCAUCCGCGCGGUGACCAUCGACCGGGUGAUGAGCUACCUCAACGCCAGU 318 hIL12AB_029AUGUGCCACCAACAGCUGGUGAUCAGCUGGUUCAGCCUGGUGUUUCUGGCCUCCCC ORFUCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGCUGGACUGGUACCCUGACGCCCCCGGCGAAAUGGUGGUGCUGACGUGCGACACCCCCGAGGAGGAUGGCAUCACCUGGACCCUGGACCAAAGCAGCGAGGUCCUCGGAAGCGGCAAGACCCUCACUAUCCAAGUGAAGGAGUUCGGGGAUGCGGGCCAGUACACCUGCCACAAGGGCGGCGAGGUGCUGUCUCAUAGCCUGCUGCUCCUGCAUAAGAAGGAAGACGGCAUCUGGAGCACCGACAUACUGAAGGAUCAGAAGGAGCCCAAGAACAAGACCUUCCUGAGGUGCGAGGCCAAGAACUACUCCGGGCGCUUCACCUGUUGGUGGCUGACCACCAUCUCCACCGACCUGACCUUCAGCGUGAAGAGCAGCAGGGGGAGCAGCGACCCCCAGGGGGUGACCUGCGGAGCCGCGACCUUGUCGGCCGAGCGGGUGAGGGGCGACAAUAAGGAGUACGAGUACUCGGUCGAAUGCCAGGAGGACUCCGCCUGCCCCGCCGCCGAGGAGUCCCUCCCCAUCGAAGUGAUGGUGGACGCCGUCCACAAGCUGAAGUACGAGAACUACACCAGCAGCUUCUUCAUACGGGAUAUCAUCAAGCCCGACCCCCCGAAGAACCUGCAGCUGAAACCCUUGAAGAACUCCAGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGUCCACCCCGCACUCAUACUUCAGCCUGACCUUCUGUGUACAGGUCCAGGGCAAGAGCAAGAGGGAAAAGAAGGAUAGGGUGUUCACCGACAAGACCUCCGCCACGGUGAUCUGUCGGAAAAACGCCAGCAUCUCCGUGCGGGCCCAGGACAGGUACUAUUCCAGCAGCUGGAGCGAGUGGGCCUCCGUCCCCUGCUCCGGCGGCGGUGGCGGGGGCAGCAGGAACCUCCCCGUGGCCACCCCCGAUCCCGGGAUGUUCCCAUGCCUGCACCACAGCCAAAACCUGCUGAGGGCCGUCUCCAAUAUGCUGCAGAAGGCGAGGCAGACCCUGGAGUUCUACCCCUGUACCUCCGAGGAGAUCGACCACGAGGAUAUCACCAAGGACAAGACCUCCACGGUCGAGGCGUGCCUGCCCCUGGAGCUCACGAAGAACGAGAGCUGCCUUAACUCCAGGGAAACCUCGUUUAUCACGAACGGCAGCUGCCUGGCGUCACGGAAGACCUCCUUUAUGAUGGCCCUAUGUCUGUCCUCGAUCUACGAGGACCUGAAGAUGUACGAGGUGGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGAUCCCAAGAGGCAGAUUUUCCUGGACCAGAACAUGCUGGCCGUGAUUGACGAGCUGAUGCAGGCGCUGAACUUCAACAGCGAGACAGUGCCGCAGAAGAGCUCCCUGGAGGAGCCGGACUUUUACAAGACCAAGAUAAAGCUGUGCAUCCUGCUCCACGCCUUCAGAAUACGGGCCGUCACCAUCGAUAGGGUGAUGUCUUACCUGAACGCCUCC 319 hIL12AB_030AUGUGCCACCAGCAGCUGGUGAUUAGCUGGUUUAGCCUGGUGUUCCUGGCAAGCCC ORFCCUGGUGGCCAUCUGGGAACUGAAAAAGGACGUGUACGUGGUCGAGCUGGAUUGGUACCCCGACGCCCCCGGCGAAAUGGUGGUGCUGACGUGUGAUACCCCCGAGGAGGACGGGAUCACCUGGACCCUGGAUCAGAGCAGCGAGGUGCUGGGGAGCGGGAAGACCCUGACGAUCCAGGUCAAGGAGUUCGGCGACGCUGGGCAGUACACCUGUCACAAGGGCGGGGAGGUGCUGUCCCACUCCCUGCUGCUCCUGCAUAAGAAAGAGGACGGCAUCUGGUCCACCGACAUCCUCAAGGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGCGGUGUGAGGCGAAGAACUACAGCGGCCGUUUCACCUGCUGGUGGCUGACGACAAUCAGCACCGACUUGACGUUCUCCGUGAAGUCCUCCAGAGGCAGCUCCGACCCCCAAGGGGUGACGUGCGGCGCGGCCACCCUGAGCGCCGAGCGGGUGCGGGGGGACAACAAGGAGUACGAGUACUCCGUGGAGUGCCAGGAGGACAGCGCCUGUCCCGCAGCCGAGGAGUCCCUGCCCAUCGAAGUCAUGGUGGACGCCGUCCACAAGCUGAAGUACGAGAACUACACCAGCAGCUUCUUCAUCCGCGAUAUCAUCAAGCCCGAUCCCCCCAAAAACCUGCAACUGAAGCCGCUGAAGAAUAGCAGGCAGGUGGAGGUGUCCUGGGAGUACCCGGACACCUGGAGCACGCCCCACAGCUAUUUCAGCCUGACCUUUUGCGUGCAGGUCCAGGGGAAGAGCAAGCGGGAGAAGAAGGACCGCGUGUUUACGGACAAAACCAGCGCCACCGUGAUCUGCAGGAAGAACGCCAGCAUCAGCGUGAGGGCCCAGGACAGGUACUACAGCAGCUCCUGGAGCGAGUGGGCCUCCGUGCCCUGUUCCGGAGGCGGCGGGGGCGGUUCCCGGAACCUCCCGGUGGCCACCCCCGACCCGGGCAUGUUCCCGUGCCUGCACCACUCACAGAAUCUGCUGAGGGCCGUGAGCAAUAUGCUGCAGAAGGCAAGGCAGACCCUGGAGUUUUAUCCCUGCACCAGCGAGGAGAUCGAGGAGGAAGACAUGAGCAAGGAGAAGACCAGCACAGUGGAGGCCUGCCUGCCCCUGGAACUGACCAAGAACGAGUCCUGUCUGAACUCCCGGGAAACCAGCUUCAUAACCAACGGCUCCUGUCUCGCCAGCAGGAAGACCAGCUUCAUGAUGGCCCUGUGCCUCAGCUCCAUCUACGAGGACCUCAAGAUGUACCAGGUUGAGUUCAAGACCAUGAACGCCAAGCUCCUGAUGGACCCCAAGAGGCAGAUCUUCCUGGACCAGAAUAUGCUGGCCGUGAUCGAUGAGUUAAUGCAGGCGCUGAACUUCAACAGCGAGACGGUGCCCCAAAAGUCCUCGCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUCCUGCACGCCUUCCGAAUCCGGGCCGUAACCAUCGACAGGGUGAUGAGCUAUCUCAACGCCUCC 320 hIL12AB_031AUGUGCCACCAGCAGCUCGUGAUCAGCUGGUUCUCGCUUGUGUUCCUGGCCUCCCC ORFCCUCGUCGCCAUCUGGGAGCUGAAGAAAGACGUGUACGUGGUGGAGCUGGACUGGUAUCCCGACGCCCCGGGGGAGAUGGUGGUGCUGACCUGCGACACCCCGGAAGAGGACGGCAUCACCUGGACGCUCGACCAGUCGUCCGAAGUGCUGGGGUCGGGCAAGACCCUCACCAUCCAGGUGAAGGAGUUCGGAGACGCCGGCCAGUACACCUGUCAUAAGGGGGGGGAGGUGCUGAGCCACAGCCUCCUGCUCCUGCACAAAAAGGAGGACGGCAUCUGGAGCACCGAUAUCCUCAAGGACCAGAAGGAGCCCAAGAACAAGACGUUCCUGAGGUGUGAGGCCAAGAACUACAGCGGGCGGUUCACGUGUUGGUGGCUCACCACCAUCUCCACCGACCUCACCUUCUCCGUGAAGUCAAGCAGGGGCAGCUCCGACCCCCAAGGCGUCACCUGCGGCGCCGCCACCCUGAGCGCCGAGAGGGUCAGGGGGGAUAACAAGGAAUACGAGUACAGUGUGGAGUGCCAAGAGGAUAGCGCCUGUCCCGCCGCCGAAGAGAGCCUGCCCAUCGAAGUGAUGGUGGACGCCGUGCACAAGCUGAAGUACGAGAACUACACCUCCAGCUUCUUCAUCAGGGAUAUCAUCAAGCCCGAUCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCAGGCAGGUGGAGGUGAGCUGGGAGUAUCCCGACACGUGGAGCACCCCGCACAGCUACUUCUCGCUGACCUUCUGCGUGCAGGUGCAAGGGAAGUCCAAGAGGGAGAAGAAGGAUAGGGUGUUCACCGACAAAACGAGCGCCACCGUGAUCUGCCGGAAGAAUGCCAGCAUCUCUGUGAGGGCCCAGGACAGGUACUAUUCCAGCUCCUGGUCGGAGUGGGCCAGCGUGCCCUGUAGCGGCGGGGGCGGGGGCGGCAGCAGGAACCUCCCGGUUGCCACCCCCGACCCCGGCAUGUUUCCGUGCCUGCACCACUCGCAAAACCUGCUGCGCGCGGUCUCCAACAUGCUGCAAAAAGCGCGCCAGACGCUGGAGUUCUACCCCUGCACCAGCGAGGAGAUCGAUCAUGAAGAUAUCACCAAAGACAAGACCUCGACCGUGGAGGCCUGCCUGCCCCUGGAGCUCACCAAGAACGAAAGCUGCCUGAACAGCAGGGAGACAAGCUUCAUCACCAACGGCAGCUGCCUGGCCUCCCGGAAGACCAGCUUCAUGAUGGCCCUGUGCCUGUCCAGCAUCUACGAGGAUCUGAAGAUGUACCAAGUGGAGUUUAAGACCAUGAACGCCAAGCUGUUAAUGGACCCCAAAAGGCAGAUCUUCCUGGAUCAGAACAUGCUGGCCGUCAUCGACGAGCUGAUGCAAGCCCUGAACUUCAACAGCGAGACGGUGCCCCAGAAGAGCAGCCUCGAGGAGCCCGACUUCUAUAAGACCAAGAUAAAGCUGUGCAUUCUGCUGCACGCCUUCAGAAUCAGGGCCGUGACCAUCGAUAGGGUGAUGAGCUACCUGAACGCCAGC 321 hIL12AB_032AUGUGUCACCAGCAGCUGGUGAUUUCCUGGUUCAGUCUGGUGUUUCUUGCCAGCCC ORFCCUGGUGGCCAUCUGGGAGCUGAAGAAAGACGUAUACGUCGUGGAGCUGGACUGGUAUCCCGACGCUCCCGGCGAGAUGGUGGUCCUCACCUGCGACACCCCAGAGGAGGACGGCAUCACCUGGACCCUGGACCAGAGCUCCGAGGUCCUGGGCAGCGGUAAGACCCUCACCAUCCAGGUGAAGGAGUUUGGUGAUGCCGGGCAGUAUACCUGCCACAAGGGCGGCGAGGUGCUGUCCCACAGCCUCCUGUUACUGCAUAAGAAGGAGGAUGGCAUCUGGAGCACCGACAUCCUCAAGGACCAGAAAGAGCCCAAGAACAAGACCUUUCUGCGGUGCGAGGCGAAAAAUUACUCCGGCCGGUUCACCUGCUGGUGGCUGACCACCAUCAGCACGGACCUGACGUUCUCCGUGAAGUCGAGCAGGGGGAGCUCCGAUCCCCAGGGCGUGACCUGCGGCGCGGCCACCCUGAGCGCCGAGCGCGUCCGCGGGGACAAUAAGGAAUACGAAUAUAGCGUGGAGUGCCAGGAGGACAGCGCCUGCCCCGCGGCCGAGGAGAGCCUCCCGAUCGAGGUGAUGGUGGAUGCCGUCCACAAGCUCAAAUACGAAAACUACACCAGCAGCUUCUUCAUUAGGGACAUCAUCAAGCCCGACCCCCCCAAAAACCUGCAGCUGAAGCCCCUGAAGAACAGCCGCCAGGUCGAGGUGUCAUGGGAGUACCCAGACACCUGGAGCACCCCCCACUCCUACUUCAGCCUGACCUUCUGCGUCCAGGUGCAGGGAAAGUCCAAACGGGAGAAGAAGGAUAGGGUCUUUACCGAUAAGACGUCGGCCACCGUCAUCUGCAGGAAGAACGCCAGCAUAAGCGUGCGGGCGCAGGAUCGGUACUACAGCUCGAGCUGGUCCGAAUGGGCCUCCGUGCCCUGUAGCGGAGGGGGUGGCGGGGGCAGCAGGAACCUGCCCGUGGCCACCCCGGACCCGGGCAUGUUUCCCUGCCUGCAUCACAGUCAGAACCUGCUGAGGGCCGUGAGCAACAUGCUCCAGAAGGCCCGCCAGACCCUGGAGUUUUACCCCUGCACCAGCGAAGAGAUCGAUCACGAAGACAUCACCAAAGACAAGACCUCCACCGUGGAGGCCUGUCUGCCCCUGGAGCUGACCAAGAACGAGAGCUGUCUGAACAGCAGGGAGACCUCCUUCAUCACCAACGGCUCCUGCCUGGCAUCCCGGAAGACCAGCUUCAUGAUGGCCCUGUGUCUGAGCUCUAUCUACGAGGACCUGAAGAUGUACCAGGUCGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGACAGAUAUUCCUGGACCAGAACAUGCUCGCCGUGAUCGAUGAACUGAUGCAAGCCCUGAACUUCAAUAGCGAGACCGUGCCCCAGAAAAGCAGCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAACUGUGCAUACUGCUGCACGCGUUCAGGAUCCGGGCCGUCACCAUCGACCGGGUGAUGUCCUAUCUGAAUGCCAGC 322 hIL12AB_033AUGUGCCACCAGCAGCUCGUGAUUAGCUGGUUUUCGCUGGUGUUCCUGGCCAGCCC ORFUCUCGUGGCCAUCUGGGAGCUGAAAAAAGACGUGUACGUGGUGGAGCUGGACUGGUACCCGGACGCCCCCGGCGAGAUGGUGGUGCUGACGUGCGACACCCCGGAAGAGGACGGCAUCACCUGGACCCUGGACCAGUCAUCCGAGGUCCUGGGCAGCGGCAAGACGCUCACCAUCCAGGUGAAGGAGUUCGGCGACGCCGGCCAGUACACAUGCCAUAAGGGCGGGGAGGUGCUGAGCCACAGCCUGCUCCUCCUGCACAAGAAGGAGGAUGGCAUCUGGUCUACAGACAUCCUGAAGGACCAGAAAGAGCCCAAGAACAAGACCUUCCUCCGGUGCGAGGCCAAGAACUACUCCGGGCGGUUUACUUGUUGGUGGCUGACCACCAUCAGCACCGACCUCACCUUCAGCGUGAAGAGCUCCCGAGGGAGCUCCGACCCCCAGGGGGUCACCUGCGGCGCCGCCACCCUGAGCGCCGAGCGGGUGAGGGGCGACAACAAGGAGUAUGAAUACAGCGUGGAAUGCCAAGAGGACAGCGCCUGUCCCGCGGCCGAGGAAAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUCCACAAACUCAAGUACGAGAACUACACCAGCAGUUUCUUCAUUCGCGACAUCAUCAAGCCGGACCCCCCCAAAAACCUGCAGCUCAAACCCCUGAAGAACAGCAGGCAGGUGGAGGUCAGCUGGGAGUACCCGGACACCUGGAGCACCCCCCAUAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGCAAGAGCAAACGCGAGAAGAAGGACCGGGUGUUUACCGACAAGACCAGCGCCACGGUGAUCUGCCGAAAGAAUGCAAGCAUCUCCGUGAGGGCGCAGGACCGCUACUACUCUAGCAGCUGGAGCGAGUGGGCCAGCGUGCCCUGCAGCGGUGGCGGCGGAGGCGGCAGCCGUAACCUCCCCGUGGCCACCCCCGACCCCGGCAUGUUCCCGUGUCUGCACCACUCCCAGAACCUGCUGAGGGCCGUCAGCAAUAUGCUGCAGAAGGCCCGGCAGACGCUGGAGUUCUACCCCUGCACCUCCGAGGAGAUCGACCAUGAGGACAUUACCAAGGACAAGACGAGCACUGUGGAGGCCUGCCUGCCCCUGGAGCUCACCAAAAACGAGAGCUGCCUGAAUAGCAGGGAGACGUCCUUCAUCACCAACGGCAGCUGUCUGGCCAGCAGGAAGACCAGCUUCAUGAUGGCCCUGUGCCUCUCCUCCAUAUAUGAGGAUCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGAUCCCAAGAGGCAGAUCUUCCUGGACCAGAAUAUGCUGGCCGUGAUUGACGAGCUGAUGCAGGCCCUGAACUUUAAUAGCGAGACCGUCCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUCUAUAAGACCAAGAUCAAGCUGUGCAUACUGCUGCACGCGUUUAGGAUAAGGGCCGUCACCAUCGACAGGGUGAUGAGCUACCUGAAUGCCAGC 323 hIL12AB_034AUGUGCCACCAACAGCUGGUGAUCUCCUGGUUCAGCCUGGUGUUCCUCGCCAGCCC ORFCCUGGUGGCCAUCUGGGAGCUGAAGAAAGACGUGUACGUGGUGGAGCUGGACUGGUAUCCCGACGCCCCCGGCGAGAUGGUCGUGCUGACCUGCGACACCCCGGAGGAGGACGGCAUCACCUGGACCCUGGAUCAGUCCUCCGAGGUGCUGGGCAGCGGGAAGACCCUGACCAUCCAGGUGAAAGAGUUCGGAGAUGCCGGCCAGUAUACCUGUCACAAGGGGGGUGAGGUGCUGAGCCAUAGCCUCUUGCUUCUGCACAAGAAGGAGGACGGCAUCUGGUCCACCGACAUCCUCAAGGACCAAAAGGAGCCGAAGAAUAAAACGUUCCUGAGGUGCGAAGCCAAGAACUAUUCCGGACGGUUCACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUCACCUUCUCCGUAAAGUCAAGCAGGGGCAGCUCCGACCCCCAGGGCGUGACCUGCGGAGCCGCCACCCUGAGCGCAGAGAGGGUGAGGGGCGACAACAAGGAGUACGAAUACUCCGUCGAGUGCCAGGAGGACAGCGCCUGCCCCGCCGCCGAGGAAAGUCUGCCCAUCGAGGUGAUGGUGGACGCCGUGCACAAGCUCAAAUACGAGAACUACACCAGCAGCUUCUUCAUCCGGGAUAUCAUCAAGCCCGACCCUCCAAAGAAUCUGCAGCUGAAACCCCUUAAGAACAGCAGGCAGGUGGAGGUCAGCUGGGAGUACCCCGACACCUGGAGCACGCCCCACUCCUACUUUAGCCUGACCUUUUGCGUGCAGGUGCAGGGGAAAAGCAAGCGGGAGAAGAAGGACAGGGUGUUCACCGAUAAGACCUCCGCUACCGUGAUCUGCAGGAAGAACGCCUCAAUCAGCGUGAGGGCCCAGGAUCGGUACUACUCCAGCUCCUGGAGCGAGUGGGCCAGCGUGCCCUGCUCUGGCGGUGGCGGCGGGGGCAGCCGGAACCUGCCGGUGGCCACUCCCGACCCGGGCAUGUUCCCGUGCCUCCACCAUUCCCAGAACCUGCUGCGGGCCGUGUCCAAUAUGCUCCAGAAGGCAAGGCAGACCCUGGAGUUCUACCCCUGCACCAGCGAGGAGAUCGAUCACGAGGACAUCACCAAAGACAAAACCAGCACGGUCGAGGCCUGCCUGCCCCUGGAACUCACCAAGAACGAAAGCUGUCUCAACAGCCGCGAGACCAGCUUCAUAACCAACGGUUCCUGUCUGGCCUCCCGCAAGACCAGCUUUAUGAUGGCCCUCUGUCUGAGCUCCAUCUAUGAAGACCUGAAAAUGUACCAGGUGGAGUUCAAAACCAUGAACGCCAAGCUUCUGAUGGACCCCAAGAGGCAGAUCUUCCUGGAUCAGAACAUGCUGGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUUAACUCCGAGACCGUGCCCCAGAAAAGCAGCCUGGAAGAGCCCGAUUUCUACAAAACGAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCCGGAUCCGUGCGGUGACCAUCGAUAGGGUGAUGAGCUACCUGAACGCCAGC 324 hIL12AB_035AUGUGCCACCAACAGCUGGUAAUCAGCUGGUUCAGCCUGGUUUUCCUCGCGUCGCC ORFCCUGGUGGCCAUCUGGGAGUUAAAGAAGGACGUGUACGUGGUGGAGCUGGAUUGGUACCCCGACGCCCCGGGCGAGAUGGUCGUGCUCACCUGCGAUACCCCCGAGGAGGACGGGAUCACCUGGACCCUGGACCAAUCCAGCGAGGUGCUGGGCAGCGGCAAGACCCUGACCAUACAGGUGAAGGAAUUUGGGGACGCCGGGCAGUACACCUGCCACAAGGGCGGGGAAGUGCUGUCCCACUCCCUCCUGCUGCUGCAUAAGAAGGAGGACGGCAUCUGGAGCACCGACAUCCUGAAGGACCAAAAGGAGCCCAAGAACAAGACCUUCCUGAGGUGCGAGGCCAAAAACUAUUCCGGCCGCUUUACCUGUUGGUGGCUGACCACCAUCUCCACCGAUCUGACCUUCAGCGUGAAGUCGUCUAGGGGCUCCUCCGACCCCCAGGGCGUAACCUGCGGCGCCGCGACCCUGAGCGCCGAGAGGGUGCGGGGCGAUAACAAAGAGUACGAGUACUCGGUGGAGUGCCAGGAGGACAGCGCCUGUCCGGCGGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUCCACAAGCUGAAGUACGAGAACUACACCAGUUCGUUCUUCAUCAGGGACAUCAUCAAGCCGGACCCCCCCAAGAACCUCCAGCUGAAGCCCCUGAAGAACAGCAGGCAGGUGGAAGUGUCCUGGGAGUAUCCCGACACCUGGAGCACCCCCCACAGCUACUUCAGCCUGACCUUUUGCGUGCAGGUGCAGGGCAAAAGCAAGAGGGAAAAGAAGGACCGGGUGUUCACCGAUAAGACGAGCGCCACCGUUAUCUGCAGGAAGAACGCCUCCAUAAGCGUGAGGGCGCAGGACCGUUACUACAGCAGCAGCUGGAGUGAGUGGGCAAGCGUGCCCUGUAGCGGCGGGGGCGGGGGCGGGUCCCGCAACCUCCCCGUCGCCACCCCCGACCCAGGCAUGUUUCCGUGCCUGCACCACAGCCAGAACCUGCUGCGGGCCGUUAGCAACAUGCUGCAGAAGGCCAGGCAGACCCUCGAGUUCUAUCCCUGCACAUCUGAGGAGAUCGACCACGAAGACAUCACUAAGGAUAAGACCUCCACCGUGGAGGCCUGUCUGCCCCUCGAGCUGACCAAGAAUGAAUCCUGCCUGAACAGCCGAGAGACCAGCUUUAUCACCAACGGCUCCUGCCUGGCCAGCAGGAAGACCUCCUUCAUGAUGGCCCUGUGCCUCUCCAGCAUCUACGAGGAUCUGAAGAUGUACCAGGUAGAGUUCAAGACGAUGAACGCCAAGCUCCUGAUGGACCCCAAGAGGCAGAUAUUCCUGGACCAGAACAUGCUGGCGGUGAUCGACGAGCUGAUGCAGGCCCUGAAUUUCAACAGCGAGACGGUGCCACAGAAGUCCAGCCUGGAGGAGCCAGACUUCUACAAGACCAAGAUCAAACUGUGCAUCCUCCUGCACGCGUUCAGGAUCCGCGCCGUCACCAUAGACAGGGUGAUGAGUUAUCUGAACGCCAGC 325 hIL12AB_036AUGUGCCAUCAGCAGCUGGUAAUCAGCUGGUUUAGCCUGGUGUUCCUGGCCAGCCC ORFACUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAACUGGACUGGUACCCCGACGCCCCUGGCGAGAUGGUGGUACUGACCUGUGACACCCCGGAGGAAGACGGUAUCACCUGGACCCUGGAUCAGAGCUCCGAGGUGCUGGGCUCCGGCAAGACACUGACCAUCCAAGUUAAGGAAUUUGGGGACGCCGGCCAGUACACCUGCCACAAGGGGGGCGAGGUGCUGUCCCACUCCCUGCUGCUUCUGCAUAAGAAGGAGGAUGGCAUCUGGUCCACCGACAUACUGAAGGACCAGAAGGAGCCCAAGAAUAAGACCUUCCUGAGAUGCGAGGCCAAGAACUACUCGGGAAGGUUCACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUGACCUUCUCCGUGAAGAGCUCCCGGGGCAGCUCCGACCCCCAGGGCGUAACCUGUGGGGCCGCUACCCUGUCCGCCGAGAGGGUCCGGGGCGACAACAAGGAAUACGAGUACAGCGUGGAGUGCCAGGAGGACUCCGCCUGCCCCGCCGCCGAGGAGUCGCUGCCCAUAGAGGUGAUGGUGGACGCCGUGCACAAGCUCAAGUACGAGAAUUACACCAGCAGCUUCUUUAUCAGGGACAUAAUUAAGCCGGACCCCCCAAAGAAUCUGCAGCUGAAGCCCCUGAAGAAUAGCCGGCAGGUGGAAGUGUCCUGGGAGUACCCCGACACCUGGAGCACCCCCCACUCCUAUUUCUCACUGACAUUCUGCGUGCAGGUGCAAGGGAAAAGCAAGAGGGAGAAGAAGGAUAGGGUGUUCACCGACAAGACAAGCGCCACCGUGAUCUGCCGAAAAAAUGCCAGCAUCAGCGUGAGGGCCCAGGAUCGGUAUUACAGCAGCUCCUGGAGCGAGUGGGCCAGCGUGCCCUGUUCCGGCGGGGGAGGGGGCGGCUCCCGGAACCUGCCGGUGGCCACCCCCGACCCUGGCAUGUUCCCCUGCCUGCAUCACAGCCAGAACCUGCUCCGGGCCGUGUCGAACAUGCUGCAGAAGGCCCGGCAGACCCUCGAGUUUUACCCCUGCACCAGCGAAGAGAUCGACCACGAAGACAUAACCAAGGACAAGACCAGCACGGUGGAGGCCUGCCUGCCCCUGGAGCUUACCAAAAACGAGUCCUGCCUGAACAGCCGGGAAACCAGCUUCAUAACGAACGGGAGCUGCCUGGCCUCCAGGAAGACCAGCUUCAUGAUGGCGCUGUGUCUGUCCAGCAUAUACGAGGAUCUGAAGAUGUAUCAGGUGGAAUUCAAAACUAUGAAUGCCAAGCUCCUGAUGGACCCCAAGAGGCAGAUCUUCCUGGACCAGAACAUGCUAGCCGUGAUCGACGAGCUGAUGCAGGCCCUCAACUUCAACUCGGAGACGGUGCCCCAGAAGUCCAGCCUCGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUACUGCUGCAUGCCUUCAGGAUAAGGGCGGUGACUAUCGACAGGGUCAUGUCCUACCUGAACGCCAGC 326 hIL12AB_037AUGUGCCACCAACAACUGGUGAUCAGCUGGUUCUCCCUGGUGUUCCUGGCCAGCCC ORFCCUGGUGGCCAUCUGGGAGCUCAAAAAAGACGUGUACGUGGUGGAGCUCGAUUGGUACCCAGACGCGCCGGGGGAAAUGGUGGUGCUGACCUGCGACACCCCAGAGGAGGAUGGCAUCACGUGGACGCUGGAUCAGUCCAGCGAGGUGCUGGGGAGCGGCAAGACGCUCACCAUCCAGGUGAAGGAAUUUGGCGACGCGGGCCAGUAUACCUGUCACAAGGGCGGCGAGGUGCUGAGCCACUCCCUGCUGCUGCUGCACAAGAAGGAGGAUGGGAUCUGGUCAACCGAUAUCCUGAAAGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGCGCUGCGAGGCCAAGAACUAUAGCGGCAGGUUCACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUGACCUUCAGCGUGAAAUCCUCCAGGGGCAGCAGCGACCCCCAGGGCGUGACCUGCGGUGCCGCCACGCUCUCCGCCGAGCGAGUGAGGGGUGACAACAAGGAGUACGAGUACAGCGUGGAAUGUCAGGAGGACAGCGCCUGUCCCGCCGCCGAGGAGUCGCUGCCCAUCGAGGUGAUGGUCGACGCGGUGCACAAGCUCAAAUACGAGAAUUACACCAGCAGCUUCUUCAUCAGGGACAUCAUCAAGCCCGACCCCCCCAAGAACCUGCAGCUGAAGCCCUUGAAGAACAGCAGGCAGGUGGAGGUGAGCUGGGAGUACCCGGACACCUGGAGCACCCCCCACUCCUACUUCAGCCUGACGUUCUGUGUGCAGGUGCAGGGGAAGUCCAAGAGGGAGAAGAAGGACCGGGUGUUCACCGACAAGACCAGCGCCACCGUGAUAUGCCGCAAGAACGCGUCCAUCAGCGUUCGCGCCCAGGACCGCUACUACAGCAGCUCCUGGUCCGAAUGGGCCAGCGUGCCCUGCAGCGGUGGAGGGGGCGGGGGCUCCAGGAAUCUGCCGGUGGCCACCCCCGACCCCGGGAUGUUCCCGUGUCUGCAUCACUCCCAGAACCUGCUGCGGGCCGUGAGCAAUAUGCUGCAGAAGGCCAGGCAGACGCUCGAGUUCUACCCCUGCACCUCCGAAGAGAUCGACCAUGAGGACAUCACCAAGGACAAGACCAGCACCGUGGAGGCCUGCCUCCCCCUGGAGCUGACCAAAAACGAGAGCUGCCUGAACUCCAGGGAGACCAGCUUUAUAACCAACGGCAGCUGCCUCGCCUCCAGGAAGACCUCGUUUAUGAUGGCCCUCUGCCUGUCCAGCAUCUACGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCGAAGUUGCUCAUGGACCCCAAGAGGCAGAUCUUCCUGGACCAGAACAUGCUCGCGGUGAUCGACGAGCUGAUGCAAGCCCUGAACUUCAACAGCGAGACCGUGCCCCAGAAGAGCAGCCUGGAAGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCCGGAUCCGGGCCGUGACCAUCGACAGGGUGAUGAGCUACCUCAACGCCUCC 327 hIL12AB_038AUGUGCCACCAGCAGCUCGUGAUCAGCUGGUUCUCCCUCGUCUUCCUGGCCUCCCC ORFGCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGCUGGACUGGUAUCCCGACGCCCCCGGCGAGAUGGUGGUGCUGACGUGCGACACACCAGAAGAGGACGGGAUCACAUGGACCCUGGAUCAGUCGUCCGAGGUGCUGGGGAGCGGCAAGACCCUCACCAUCCAAGUGAAGGAGUUCGGGGACGCCGGCCAGUACACCUGCCACAAGGGCGGGGAGGUGCUCUCCCAUAGCCUGCUCCUCCUGCACAAAAAGGAGGAUGGCAUCUGGAGCACCGACAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACAUUUCUCAGGUGUGAGGCCAAGAACUAUUCGGGCAGGUUUACCUGUUGGUGGCUCACCACCAUCUCUACCGACCUGACGUUCUCCGUCAAGUCAAGCAGGGGGAGCUCGGACCCCCAGGGGGUGACAUGUGGGGCCGCCACCCUGAGCGCGGAGCGUGUCCGCGGCGACAACAAGGAGUACGAGUAUUCCGUGGAGUGCCAGGAGGACAGCGCCUGCCCCGCCGCCGAGGAGUCCCUGCCCAUAGAGGUGAUGGUGGACGCCGUCCACAAGUUGAAGUACGAAAAUUAUACCUCCUCGUUCUUCAUUAGGGACAUCAUCAAGCCUGACCCCCCGAAGAACCUACAACUCAAGCCCCUCAAGAACUCCCGCCAGGUGGAGGUGUCCUGGGAGUACCCCGACACCUGGUCCACCCCGCACAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUCCAGGGGAAGAGCAAGCGUGAAAAGAAAGACAGGGUGUUCACCGACAAGACGAGCGCCACCGUGAUCUGCAGGAAAAACGCCUCCAUCUCCGUGCGCGCCCAGGACAGGUACUACAGUAGCUCCUGGAGCGAAUGGGCCAGCGUGCCGUGCAGCGGCGGGGGAGGAGGCGGCAGUCGCAACCUGCCCGUGGCCACCCCCGACCCCGGCAUGUUCCCAUGCCUGCACCACAGCCAGAACCUGCUGAGGGCAGUCAGCAAUAUGCUGCAGAAGGCCAGGCAGACCCUGGAGUUUUAUCCCUGCACCAGCGAGGAGAUCGACCACGAGGACAUCACCAAGGACAAGACCUCCACCGUCGAGGCCUGCCUGCCACUGGAGCUGACCAAAAACGAGAGCUGCCUGAACUCCAGGGAGACCUCCUUCAUCACCAACGGGAGCUGCCUGGCCAGCCGGAAGACCAGCUUCAUGAUGGCGCUGUGCCUCAGCAGCAUCUACGAGGAUCUCAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCGAAGCUGCUGAUGGACCCCAAGCGGCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUGAUUGACGAGCUCAUGCAGGCCCUGAACUUCAAUAGCGAGACCGUCCCCCAAAAGAGCAGCCUGGAGGAACCCGACUUCUACAAAACGAAGAUCAAGCUCUGCAUCCUGCUGCACGCCUUCCGGAUCCGGGCCGUGACCAUCGAUCGUGUGAUGAGCUACCUGAACGCCUCG 328 hIL12AB_039AUGUGCCACCAGCAGCUCGUCAUCUCCUGGUUUAGCCUGGUGUUUCUGGCCUCCCC ORFCCUGGUCGCCAUCUGGGAGCUGAAGAAAGACGUGUACGUGGUGGAGCUGGACUGGUACCCGGACGCUCCCGGGGAGAUGGUGGUGCUGACCUGCGACACCCCCGAGGAGGACGGCAUCACCUGGACCCUGGACCAGAGCUCCGAGGUGCUGGGGAGCGGCAAGACCCUGACCAUUCAGGUGAAAGAGUUCGGCGACGCCGGCCAAUAUACCUGCCACAAGGGGGGGGAGGUCCUGUCGCAUUCCCUGCUGCUGCUUCACAAAAAGGAGGAUGGCAUCUGGAGCACCGACAUCCUGAAGGACCAGAAAGAACCCAAGAACAAGACGUUCCUGCGCUGCGAGGCCAAGAACUACAGCGGCCGGUUCACCUGUUGGUGGCUGACCACCAUCUCCACCGACCUGACUUUCUCGGUGAAGAGCAGCCGCGGGAGCAGCGACCCCCAGGGAGUGACCUGCGGCGCCGCCACCCUGAGCGCCGAAAGGGUGAGGGGCGACAAUAAAGAGUACGAGUAUUCCGUGGAGUGCCAGGAGGACAGCGCCUGUCCCGCCGCCGAGGAGUCCCUGCCUAUCGAGGUGAUGGUCGACGCGGUGCACAAGCUCAAGUACGAAAACUACACCAGCAGCUUUUUCAUCAGGGAUAUCAUCAAACCAGACCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAAAACAGCAGGCAGGUGGAAGUGAGCUGGGAAUACCCCGAUACCUGGUCCACCCCCCACAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGGAAGUCCAAGCGGGAGAAGAAAGAUCGGGUGUUCACGGACAAGACCAGCGCCACCGUGAUUUGCAGGAAAAACGCCAGCAUCUCCGUGAGGGCUCAGGACAGGUACUACAGCUCCAGCUGGAGCGAGUGGGCCUCCGUGCCUUGCAGCGGGGGAGGAGGCGGCGGCAGCAGGAAUCUGCCCGUCGCAACCCCCGACCCCGGCAUGUUCCCCUGCCUGCACCACAGCCAGAAUCUGCUGCGAGCCGUGAGCAACAUGCUCCAGAAGGCCCGGCAGACGCUGGAGUUCUACCCCUGCACCUCCGAGGAGAUCGACCACGAGGACAUCACCAAGGAUAAGACGAGCACCGUCGAGGCCUGUCUCCCCCUGGAGCUCACCAAGAACGAGUCCUGCCUGAAUAGCAGGGAGACGUCCUUCAUAACCAACGGCAGCUGUCUGGCGUCCAGGAAGACCAGCUUCAUGAUGGCCCUCUGCCUGAGCUCCAUCUACGAGGACCUCAAGAUGUACCAGGUCGAGUUCAAGACCAUGAACGCAAAACUGCUCAUGGAUCCAAAGAGGCAGAUCUUUCUGGACCAGAACAUGCUGGCCGUGAUCGAUGAACUCAUGCAGGCCCUGAAUUUCAAUUCCGAGACCGUGCCCCAGAAGAGCUCCCUGGAGGAACCCGACUUCUACAAAACAAAGAUCAAGCUGUGUAUCCUCCUGCACGCCUUCCGGAUCAGGGCCGUCACCAUUGACCGGGUGAUGUCCUACCUGAACGCCAGC 329 hIL12AB_040AUGUGCCAUCAGCAGCUGGUGAUCAGCUGGUUCAGCCUCGUGUUCCUCGCCAGCCC ORFCCUCGUGGCCAUCUGGGAGCUGAAAAAGGACGUGUACGUGGUGGAGCUGGACUGGUAUCCCGACGCCCCGGGCGAGAUGGUGGUGCUGACCUGCGACACCCCCGAGGAGGACGGCAUUACCUGGACACUGGACCAGAGCAGCGAGGUCCUGGGCAGCGGGAAGACCCUGACAAUUCAGGUGAAGGAGUUCGGCGACGCCGGACAGUACACGUGCCACAAGGGGGGGGAGGUGCUGUCCCACAGCCUCCUCCUGCUGCACAAGAAGGAGGAUGGCAUCUGGAGCACCGACAUCCUGAAGGAUCAGAAGGAGCCCAAGAACAAGACCUUUCUGAGAUGCGAGGCCAAGAAUUACAGCGGCCGUUUCACCUGCUGGUGGCUCACCACCAUCAGCACCGACCUGACCUUCAGCGUGAAAUCCUCCAGGGGCUCCUCCGACCCGCAGGGAGUGACCUGCGGCGCCGCCACACUGAGCGCCGAGCGGGUCAGAGGGGACAACAAGGAGUACGAGUACAGCGUUGAGUGCCAGGAGGACAGCGCCUGUCCCGCGGCCGAGGAAUCCCUGCCCAUCGAGGUGAUGGUGGACGCAGUGCACAAGCUGAAGUACGAGAACUAUACCUCGAGCUUCUUCAUCCGGGAUAUCAUUAAGCCCGAUCCCCCGAAGAACCUGCAGCUCAAACCCCUGAAGAACAGCAGGCAGGUGGAGGUCUCCUGGGAGUACCCCGACACAUGGUCCACCCCCCAUUCCUAUUUCUCCCUGACCUUUUGCGUGCAGGUGCAGGGCAAGAGCAAGAGGGAGAAAAAGGACAGGGUGUUCACCGACAAGACCUCCGCCACCGUGAUCUGCCGUAAGAACGCUAGCAUCAGCGUCAGGGCCCAGGACAGGUACUAUAGCAGCUCCUGGUCCGAGUGGGCCAGCGUCCCGUGCAGCGGCGGGGGCGGUGGAGGCUCCCGGAACCUCCCCGUGGCCACCCCGGACCCCGGGAUGUUUCCCUGCCUGCAUCACAGCCAGAACCUGCUGAGGGCCGUGUCCAACAUGCUGCAGAAGGCCAGGCAGACACUCGAGUUUUACCCCUGCACCAGCGAGGAGAUCGACCACGAAGACAUCACCAAGGACAAGACCUCCACCGUGGAGGCAUGCCUGCCCCUGGAGCUGACCAAAAACGAAAGCUGUCUGAACUCCAGGGAGACCUCCUUUAUCACGAACGGCUCAUGCCUGGCCUCCAGAAAGACCAGCUUCAUGAUGGCCCUGUGCCUGAGCUCCAUCUACGAGGACUUGAAAAUGUACCAGGUCGAGUUCAAGACCAUGAACGCCAAGCUGCUCAUGGACCCCAAAAGGCAGAUCUUUCUGGACCAGAAUAUGCUGGCCGUGAUCGACGAGCUCAUGCAAGCCCUGAAUUUCAACAGCGAGACCGUGCCCCAGAAGUCCUCCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUACUCCUGCACGCGUUUAGGAUCAGGGCGGUGACCAUCGAUAGGGUGAUGAGCUACCUGAAUGCCUCC 330 Wild TypeAUGUGUCACCAGCAGUUGGUCAUCUCUUGGUUUUCCCUGGUUUUUCUGGCAUCUCC IL12B signalCCUCGUGGCC peptide Nucleic acids 331 Syn 5 AUUGGGCACCCGUAAGGG promoter332 hIL12AB_001 UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA(5′UTR ORF GAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGUCACCAGCAGCUGGU3′UTR) CAUUAGCUGGUUUAGCCUUGUGUUCCUGGCCUCCCCCCUUGUCGCUAUUUGGGAGCUCAAGAAGGACGUGUACGUGGUGGAGUUGGAUUGGUACCCAGACGCGCCCGGAGAGAUGGUAGUUCUGACCUGUGAUACCCCAGAGGAGGACGGCAUCACCUGGACGCUGGACCAAAGCAGCGAGGUUUUGGGCUCAGGGAAAACGCUGACCAUCCAGGUGAAGGAAUUCGGCGACGCCGGGCAGUACACCUGCCAUAAGGGAGGAGAGGUGCUGAGCCAUUCCCUUCUUCUGCUGCACAAGAAAGAGGACGGCAUCUGGUCUACCGACAUCCUGAAAGACCAGAAGGAGCCCAAGAACAAAACCUUCCUGAGGUGCGAGGCCAAGAACUACUCCGGCAGGUUCACUUGUUGGUGGCUGACCACCAUCAGUACAGACCUGACUUUUAGUGUAAAAAGCUCCAGAGGCUCGUCCGAUCCCCAAGGGGUGACCUGCGGCGCAGCCACUCUGAGCGCUGAGCGCGUGCGCGGUGACAAUAAAGAGUACGAGUACAGCGUUGAGUGUCAAGAAGAUAGCGCUUGCCCUGCCGCCGAGGAGAGCCUGCCUAUCGAGGUGAUGGUUGACGCAGUGCACAAGCUUAAGUACGAGAAUUACACCAGCUCAUUCUUCAUUAGAGAUAUAAUCAAGCCUGACCCACCCAAGAACCUGCAGCUGAAGCCACUGAAAAACUCACGGCAGGUCGAAGUGAGCUGGGAGUACCCCGACACCUGGAGCACUCCUCAUUCCUAUUUCUCUCUUACAUUCUGCGUCCAGGUGCAGGGCAAGAGCAAGCGGGAAAAGAAGGAUCGAGUCUUCACCGACAAAACAAGCGCGACCGUGAUUUGCAGGAAGAACGCCAGCAUCUCCGUCAGAGCCCAGGAUAGAUACUAUAGUAGCAGCUGGAGCGAGUGGGCAAGCGUGCCCUGUUCCGGCGGCGGGGGCGGGGGCAGCCGAAACUUGCCUGUCGCUACCCCGGACCCUGGAAUGUUUCCGUGUCUGCACCACAGCCAGAACCUGCUGAGAGCCGUGUCGAAUAUGCUCCAGAAGGCCCGGCAGACCCUUGAGUUCUACCCCUGUACCAGCGAAGAGAUCGAUCAUGAAGAUAUCACGAAAGAUAAAACAUCCACCGUCGAGGCUUGUCUCCCGCUGGAGCUGACCAAGAACGAGAGCUGUCUGAAUAGCCGGGAGACGUCUUUCAUCACGAAUGGUAGCUGUCUGGCCAGCAGGAAAACUUCCUUCAUGAUGGCUCUCUGCCUGAGCUCUAUCUAUGAAGAUCUGAAGAUGUAUCAGGUGGAGUUUAAAACAAUGAACGCCAAACUCCUGAUGGACCCAAAAAGGCAAAUCUUUCUGGACCAGAAUAUGCUGGCCGUGAUAGACGAGCUGAUGCAGGCACUGAACUUCAACAGCGAGACGGUGCCACAGAAAUCCAGCCUGGAGGAGCCUGACUUUUACAAAACUAAGAUCAAGCUGUGUAUCCUGCUGCACGCCUUUAGAAUCCGUGCCGUGACUAUCGACAGGGUGAUGUCAUACCUCAACGCUUCAUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 333 hIL12AB_002UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAGCAGCUGGU 3′UTR)GAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCAGCCCCCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGUUGGAUUGGUACCCCGACGCCCCCGGCGAGAUGGUGGUGCUGACCUGCGACACCCCCGAGGAGGACGGCAUCACCUGGACCCUGGACCAGAGCAGCGAGGUGCUGGGCAGCGGCAAGACCCUGACCAUCCAGGUGAAGGAGUUCGGCGACGCCGGCCAGUACACCUGCCACAAGGGCGGCGAGGUGCUGAGCCACAGCCUGCUGCUGCUGCACAAGAAGGAGGACGGCAUCUGGAGCACCGACAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGAGAUGCGAGGCCAAGAACUACAGCGGCAGAUUCACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUGACCUUCAGCGUGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGUGACCUGCGGCGCCGCCACCCUGAGCGCCGAGAGAGUGAGAGGCGACAACAAGGAGUACGAGUACAGCGUGGAGUGCCAGGAAGAUAGCGCCUGCCCCGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUGCACAAGCUGAAGUACGAGAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAGCCCGACCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCCGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGAGCACCCCCCACAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGCAAGAGCAAGAGAGAGAAGAAAGAUAGAGUGUUCACCGACAAGACCAGCGCCACCGUGAUCUGCAGAAAGAACGCCAGCAUCAGCGUGAGAGCCCAAGAUAGAUACUACAGCAGCAGCUGGAGCGAGUGGGCCAGCGUGCCCUGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCUGCCCGUGGCCACCCCCGACCCCGGCAUGUUCCCCUGCCUGCACCACAGCCAGAACCUGCUGAGAGCCGUGAGCAACAUGCUGCAGAAGGCCCGGCAGACCCUGGAGUUCUACCCCUGCACCAGCGAGGAGAUCGACCACGAAGAUAUCACCAAAGAUAAGACCAGCACCGUGGAGGCCUGCCUGCCCCUGGAGCUGACCAAGAACGAGAGCUGCCUGAACAGCAGAGAGACCAGCUUCAUCACCAACGGCAGCUGCCUGGCCAGCAGAAAGACCAGCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGGCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAGACCGUGCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCAGAAUCAGAGCCGUGACCAUCGACAGAGUGAUGAGCUACCUGAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 334 hIL12AB_003UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGUCACCAGCAGUUGGU 3′UTR)CAUCUCUUGGUUUUCCCUGGUUUUUCUGGCAUCUCCCCUCGUGGCCAUCUGGGAACUGAAGAAAGACGUUUACGUUGUAGAAUUGGAUUGGUAUCCGGACGCUCCUGGAGAAAUGGUGGUCCUCACCUGUGACACCCCUGAAGAAGACGGAAUCACCUGGACCUUGGACCAGAGCAGUGAGGUCUUAGGCUCUGGCAAAACCCUGACCAUCCAAGUCAAAGAGUUUGGAGAUGCUGGCCAGUACACCUGUCACAAAGGAGGCGAGGUUCUAAGCCAUUCGCUCCUGCUGCUUCACAAAAAGGAAGAUGGAAUUUGGUCCACUGAUAUUUUAAAGGACCAGAAAGAACCCAAAAAUAAGACCUUUCUAAGAUGCGAGGCCAAGAAUUAUUCUGGACGUUUCACCUGCUGGUGGCUGACGACAAUCAGUACUGAUUUGACAUUCAGUGUCAAAAGCAGCAGAGGCUCUUCUGACCCCCAAGGGGUGACGUGCGGAGCUGCUACACUCUCUGCAGAGAGAGUCAGAGGUGACAACAAGGAGUAUGAGUACUCAGUGGAGUGCCAGGAAGAUAGUGCCUGCCCAGCUGCUGAGGAGAGUCUGCCCAUUGAGGUCAUGGUGGAUGCCGUUCACAAGCUCAAGUAUGAAAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAACCUGACCCACCCAAGAACUUGCAGCUGAAGCCAUUAAAGAAUUCUCGGCAGGUGGAGGUCAGCUGGGAGUACCCUGACACCUGGAGUACUCCACAUUCCUACUUCUCCCUGACAUUCUGCGUUCAGGUCCAGGGCAAGAGCAAGAGAGAAAAGAAAGAUAGAGUCUUCACAGAUAAGACCUCAGCCACGGUCAUCUGCCGCAAAAAUGCCAGCAUUAGCGUGCGGGCCCAGGACCGCUACUAUAGCUCAUCUUGGAGCGAAUGGGCAUCUGUGCCCUGCAGUGGCGGAGGGGGCGGAGGGAGCAGAAACCUCCCCGUGGCCACUCCAGACCCAGGAAUGUUCCCAUGCCUUCACCACUCCCAAAACCUGCUGAGGGCCGUCAGCAACAUGCUCCAGAAGGCCCGGCAAACUUUAGAAUUUUACCCUUGCACUUCUGAAGAGAUUGAUCAUGAAGAUAUCACAAAAGAUAAAACCAGCACAGUGGAGGCCUGUUUACCAUUGGAAUUAACCAAGAAUGAGAGUUGCCUAAAUUCCAGAGAGACCUCUUUCAUAACUAAUGGGAGUUGCCUGGCCUCCAGAAAGACCUCUUUUAUGAUGGCCCUGUGCCUUAGUAGUAUUUAUGAAGAUUUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAAUGCAAAGCUUCUGAUGGAUCCUAAGAGGCAGAUCUUUUUAGAUCAAAACAUGCUGGCAGUUAUUGAUGAGCUGAUGCAGGCCCUGAAUUUCAACAGUGAGACGGUGCCACAAAAAUCCUCCCUUGAAGAACCAGAUUUCUACAAGACCAAGAUCAAGCUCUGCAUACUUCUUCAUGCUUUCAGAAUUCGGGCAGUGACUAUUGAUAGAGUGAUGAGCUAUCUGAAUGCUUCCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 335 hIL12AB_004UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGGGCUGCCACCAGCAGCU 3′UTR)GGUCAUCAGCUGGUUCUCCCUGGUCUUCCUGGCCAGCCCCCUGGUGGCCAUCUGGGAGCUGAAGAAAGACGUCUACGUAGUAGAGUUGGAUUGGUACCCAGACGCACCUGGAGAAAUGGUGGUUCUCACCUGUGACACGCCAGAAGAAGACGGUAUCACCUGGACGCUGGACCAGAGCUCAGAAGUUCUUGGCAGUGGAAAAACGCUGACCAUACAAGUAAAAGAAUUUGGGGAUGCUGGCCAGUACACCUGCCACAAAGGAGGAGAAGUUCUCAGCCACAGCCUGCUGCUGCUGCACAAGAAAGAAGAUGGCAUCUGGAGCACAGAUAUUUUAAAAGACCAGAAGGAGCCCAAGAACAAAACCUUCCUUCGAUGUGAGGCCAAGAACUACAGUGGCCGCUUCACCUGCUGGUGGCUCACCACCAUCAGCACAGACCUCACCUUCUCGGUGAAGAGCAGCCGUGGCAGCUCAGACCCCCAAGGAGUCACCUGUGGGGCGGCCACGCUGUCGGCAGAAAGAGUUCGAGGUGACAACAAGGAAUAUGAAUACUCGGUGGAAUGUCAAGAAGAUUCGGCCUGCCCGGCGGCAGAAGAAAGUCUUCCCAUAGAAGUCAUGGUGGAUGCUGUUCACAAAUUAAAAUAUGAAAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAGCCAGACCCGCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCCGGCAGGUGGAAGUUUCCUGGGAGUACCCAGAUACGUGGAGCACGCCGCACAGCUACUUCAGCCUCACCUUCUGUGUACAAGUACAAGGCAAGAGCAAGAGAGAGAAGAAAGAUCGUGUCUUCACAGAUAAAACCUCGGCGACGGUCAUCUGCAGGAAGAAUGCCUCCAUCUCGGUUCGAGCCCAGGACCGCUACUACAGCAGCAGCUGGAGUGAGUGGGCCUCGGUGCCCUGCAGUGGUGGCGGCGGCGGCGGCAGCAGAAACCUUCCUGUGGCCACGCCGGACCCUGGCAUGUUCCCGUGCCUGCACCACAGCCAAAAUUUACUUCGAGCUGUUUCUAACAUGCUGCAGAAAGCACGGCAAACUUUAGAAUUCUACCCCUGCACCUCAGAAGAAAUAGACCAUGAAGAUAUCACCAAAGAUAAAACCAGCACUGUAGAGGCCUGCCUGCCCCUGGAGCUCACCAAGAAUGAAUCCUGCCUCAACAGCAGAGAGACCAGCUUCAUCACCAAUGGCAGCUGCCUGGCCAGCAGGAAAACCAGCUUCAUGAUGGCGCUCUGCCUGAGCAGCAUCUAUGAAGAUUUGAAGAUGUACCAAGUAGAAUUUAAAACCAUGAAUGCCAAGCUGCUCAUGGACCCCAAGCGGCAGAUAUUUUUGGAUCAAAACAUGCUGGCUGUCAUUGAUGAGCUCAUGCAAGCAUUAAACUUCAACUCAGAGACGGUGCCCCAGAAGAGCAGCCUGGAGGAGCCAGAUUUCUACAAAACCAAGAUCAAGCUCUGCAUCUUAUUACAUGCCUUCCGCAUCCGGGCGGUCACCAUUGACCGUGUCAUGUCCUACUUAAAUGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 336 hIL12AB_005UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAGCAGCUGGU 3′UTR)CAUCAGCUGGUUCUCCCUGGUCUUCCUGGCCAGCCCCCUGGUGGCCAUCUGGGAGCUGAAGAAAGACGUCUACGUAGUAGAGUUGGAUUGGUACCCAGACGCACCUGGAGAAAUGGUGGUUCUCACCUGUGACACGCCAGAAGAAGACGGUAUCACCUGGACGCUGGACCAGAGCUCAGAAGUUCUUGGCAGUGGAAAAACGCUGACCAUACAAGUAAAAGAAUUUGGGGAUGCUGGCCAGUACACCUGCCACAAAGGAGGAGAAGUUCUCAGCCACAGCCUGCUGCUGCUGCACAAGAAAGAAGAUGGCAUCUGGAGCACAGAUAUUUUAAAAGACCAGAAGGAGCCCAAGAACAAAACCUUCCUUCGAUGUGAGGCCAAGAACUACAGUGGCCGCUUCACCUGCUGGUGGCUCACCACCAUCAGCACAGACCUCACCUUCUCGGUGAAGAGCAGCCGUGGCAGCUCAGACCCCCAAGGAGUCACCUGUGGGGCGGCCACGCUGUCGGCAGAAAGAGUUCGAGGUGACAACAAGGAAUAUGAAUACUCGGUGGAAUGUCAAGAAGAUUCGGCCUGCCCGGCGGCAGAAGAAAGUCUUCCCAUAGAAGUCAUGGUGGAUGCUGUUCACAAAUUAAAAUAUGAAAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAGCCAGACCCGCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCCGGCAGGUGGAAGUUUCCUGGGAGUACCCAGAUACGUGGAGCACGCCGCACAGCUACUUCAGCCUCACCUUCUGUGUACAAGUACAAGGCAAGAGCAAGAGAGAGAAGAAAGAUCGUGUCUUCACAGAUAAAACCUCGGCGACGGUCAUCUGCAGGAAGAAUGCCUCCAUCUCGGUUCGAGCCCAGGACCGCUACUACAGCAGCAGCUGGAGUGAGUGGGCCUCGGUGCCCUGCAGUGGUGGCGGCGGCGGCGGCAGCAGAAACCUUCCUGUGGCCACGCCGGACCCUGGCAUGUUCCCGUGCCUGCACCACAGCCAAAAUUUACUUCGAGCUGUUUCUAACAUGCUGCAGAAAGCACGGCAAACUUUAGAAUUCUACCCCUGCACCUCAGAAGAAAUAGACCAUGAAGAUAUCACCAAAGAUAAAACCAGCACUGUAGAGGCCUGCCUGCCCCUGGAGCUCACCAAGAAUGAAUCCUGCCUCAACAGCAGAGAGACCAGCUUCAUCACCAAUGGCAGCUGCCUGGCCAGCAGGAAAACCAGCUUCAUGAUGGCGCUCUGCCUGAGCAGCAUCUAUGAAGAUUUGAAGAUGUACCAAGUAGAAUUUAAAACCAUGAAUGCCAAGCUGCUCAUGGACCCCAAGCGGCAGAUAUUUUUGGAUCAAAACAUGCUGGCUGUCAUUGAUGAGCUCAUGCAAGCAUUAAACUUCAACUCAGAGACGGUGCCCCAGAAGAGCAGCCUGGAGGAGCCAGAUUUCUACAAAACCAAGAUCAAGCUCUGCAUCUUAUUACAUGCCUUCCGCAUCCGGGCGGUCACCAUUGACCGUGUCAUGUCCUACUUAAAUGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 337 hIL12AB_006UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAGCAGCUGGU 3′UTR)GAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCAGCCCCCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGUUGGAUUGGUACCCCGACGCCCCCGGCGAGAUGGUGGUGCUGACCUGUGACACCCCCGAGGAGGACGGCAUCACCUGGACCCUGGACCAGAGCAGCGAGGUGCUGGGCAGCGGCAAGACCCUGACCAUCCAGGUGAAGGAGUUCGGGGACGCCGGCCAGUACACCUGCCACAAGGGCGGCGAGGUGCUGAGCCACAGCCUGCUGCUGCUGCACAAGAAGGAGGACGGCAUCUGGAGCACAGAUAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGAGAUGCGAGGCCAAGAACUACAGCGGCAGAUUCACCUGCUGGUGGCUGACCACCAUCAGCACAGAUUUGACCUUCAGCGUGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGUGACCUGCGGCGCCGCCACCCUGAGCGCCGAGAGAGUGAGAGGUGACAACAAGGAGUACGAGUACAGCGUGGAGUGCCAGGAAGAUAGCGCCUGCCCCGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUGCACAAGCUGAAGUACGAGAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAGCCCGACCCGCCGAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCCGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGAGCACCCCCCACAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGCAAGAGCAAGAGAGAGAAGAAAGAUAGAGUGUUCACAGAUAAGACCAGCGCCACCGUGAUCUGCAGAAAGAACGCCAGCAUCAGCGUGAGAGCCCAAGAUAGAUACUACAGCAGCAGCUGGAGCGAGUGGGCCAGCGUGCCCUGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCUGCCCGUGGCCACCCCCGACCCCGGCAUGUUCCCCUGCCUGCACCACAGCCAGAACCUGCUGAGAGCCGUGAGCAACAUGCUGCAGAAGGCCCGGCAGACCCUGGAGUUCUACCCCUGCACCAGCGAGGAGAUCGACCACGAAGAUAUCACCAAAGAUAAGACCAGCACCGUGGAGGCCUGCCUGCCCCUGGAGCUGACCAAGAAUGAAAGCUGCCUGAACAGCAGAGAGACCAGCUUCAUCACCAACGGCAGCUGCCUGGCCAGCAGAAAGACCAGCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGGCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAGACCGUGCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCAGAAUCAGAGCCGUGACCAUCGACAGAGUGAUGAGCUACCUGAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 338 hIL12AB_007UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAGCAGCUUGU 3′UTR)CAUCUCCUGGUUCUCUCUUGUCUUCCUUGCUUCUCCUCUUGUGGCCAUCUGGGAGCUGAAGAAGGACGUUUACGUAGUGGAGUUGGAUUGGUACCCUGACGCACCUGGAGAAAUGGUGGUUCUCACCUGUGACACUCCUGAGGAGGACGGUAUCACCUGGACGUUGGACCAGUCUUCUGAGGUUCUUGGCAGUGGAAAAACUCUUACUAUUCAGGUGAAGGAGUUUGGAGAUGCUGGCCAGUACACCUGCCACAAGGGUGGUGAAGUUCUCAGCCACAGUUUACUUCUUCUUCACAAGAAGGAGGAUGGCAUCUGGUCUACUGACAUUUUAAAAGACCAGAAGGAGCCCAAGAACAAAACAUUCCUUCGUUGUGAAGCCAAGAACUACAGUGGUCGUUUCACCUGCUGGUGGCUUACUACUAUUUCUACUGACCUUACUUUCUCUGUGAAGUCUUCUCGUGGCUCUUCUGACCCUCAGGGUGUCACCUGUGGGGCUGCUACUCUUUCUGCUGAGCGUGUGCGUGGUGACAACAAGGAGUAUGAAUACUCGGUGGAGUGCCAGGAAGAUUCUGCCUGCCCUGCUGCUGAGGAGUCUCUUCCUAUUGAGGUGAUGGUGGAUGCUGUGCACAAGUUAAAAUAUGAAAACUACACUUCUUCUUUCUUCAUUCGUGACAUUAUAAAACCUGACCCUCCCAAGAACCUUCAGUUAAAACCUUUAAAAAACUCUCGUCAGGUGGAGGUGUCCUGGGAGUACCCUGACACGUGGUCUACUCCUCACUCCUACUUCUCUCUUACUUUCUGUGUCCAGGUGCAGGGCAAGUCCAAGCGUGAGAAGAAGGACCGUGUCUUCACUGACAAAACAUCUGCUACUGUCAUCUGCAGGAAGAAUGCAUCCAUCUCUGUGCGUGCUCAGGACCGUUACUACAGCUCUUCCUGGUCUGAGUGGGCUUCUGUGCCCUGCUCUGGCGGCGGCGGCGGCGGCAGCAGAAAUCUUCCUGUGGCUACUCCUGACCCUGGCAUGUUCCCCUGCCUUCACCACUCGCAGAACCUUCUUCGUGCUGUGAGCAACAUGCUUCAGAAGGCUCGUCAAACUUUAGAAUUCUACCCCUGCACUUCUGAGGAGAUUGACCAUGAAGAUAUCACCAAAGAUAAAACAUCUACUGUGGAGGCCUGCCUUCCUUUAGAGCUGACCAAGAAUGAAUCCUGCUUAAAUUCUCGUGAGACGUCUUUCAUCACCAAUGGCAGCUGCCUUGCCUCGCGCAAAACAUCUUUCAUGAUGGCUCUUUGCCUUUCUUCCAUCUAUGAAGAUUUAAAAAUGUACCAGGUGGAGUUCAAGACCAUGAAUGCAAAGCUUCUCAUGGACCCCAAGCGUCAGAUAUUUUUGGACCAGAACAUGCUUGCUGUCAUUGAUGAGCUCAUGCAGGCUUUAAACUUCAACUCUGAGACGGUGCCUCAGAAGUCUUCUUUAGAAGAGCCUGACUUCUACAAGACCAAGAUAAAACUUUGCAUUCUUCUUCAUGCUUUCCGCAUCCGUGCUGUGACUAUUGACCGUGUGAUGUCCUACUUAAAUGCUUCUUGAUAAUAGGCUGGAGCCUCGGUGGCCAAGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 339 hIL12AB_008UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGUCAUCAACAACUCGU 3′UTR)GAUUAGCUGGUUCAGUCUCGUGUUCCUGGCCUCUCCGCUGGUGGCCAUCUGGGAGCUUAAGAAGGACGUGUACGUGGUGGAGCUCGAUUGGUACCCCGACGCACCUGGCGAGAUGGUGGUGCUAACCUGCGAUACCCCCGAGGAGGACGGGAUCACUUGGACCCUGGAUCAGAGUAGCGAAGUCCUGGGCUCUGGCAAAACACUCACAAUCCAGGUGAAGGAAUUCGGAGACGCUGGUCAGUACACUUGCCACAAGGGGGGUGAAGUGCUGUCUCACAGCCUGCUGUUACUGCACAAGAAGGAGGAUGGGAUCUGGUCAACCGACAUCCUGAAGGAUCAGAAGGAGCCUAAGAACAAGACCUUUCUGAGGUGUGAAGCUAAGAACUAUUCCGGAAGAUUCACUUGCUGGUGGUUGACCACAAUCAGCACUGACCUGACCUUUUCCGUGAAGUCCAGCAGAGGAAGCAGCGAUCCUCAGGGCGUAACGUGCGGCGCGGCUACCCUGUCAGCUGAGCGGGUUAGAGGCGACAACAAAGAGUAUGAGUACUCCGUGGAGUGUCAGGAAGAUAGCGCCUGCCCCGCAGCCGAGGAGAGUCUGCCCAUCGAGGUGAUGGUGGACGCUGUCCAUAAGUUAAAAUACGAAAAUUACACAAGUUCCUUUUUCAUCCGCGAUAUUAUCAAACCCGAUCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAAUAGCCGACAGGUGGAAGUCUCUUGGGAGUAUCCUGACACCUGGUCCACGCCUCACAGCUACUUUAGUCUGACUUUCUGUGUCCAGGUCCAGGGCAAGAGCAAGAGAGAGAAAAAGGAUAGAGUGUUUACUGACAAAACAUCUGCUACAGUCAUCUGCAGAAAGAACGCCAGUAUCUCAGUGAGGGCGCAAGAUAGAUACUACAGUAGUAGCUGGAGCGAAUGGGCUAGCGUGCCCUGUUCAGGGGGCGGCGGAGGGGGCUCCAGGAAUCUGCCCGUGGCCACCCCCGACCCUGGGAUGUUCCCUUGCCUCCAUCACUCACAGAACCUGCUCAGAGCAGUGAGCAACAUGCUCCAAAAGGCCCGCCAGACCCUGGAGUUUUACCCUUGUACUUCAGAAGAGAUCGAUCACGAAGAUAUAACAAAGGAUAAAACCAGCACCGUGGAGGCCUGUCUGCCUCUGGAACUCACAAAGAAUGAAAGCUGUCUGAAUUCCAGGGAAACCUCCUUCAUUACUAACGGAAGCUGUCUCGCAUCUCGCAAAACAUCAUUCAUGAUGGCCCUCUGCCUGUCUUCUAUCUAUGAAGAUCUCAAGAUGUAUCAGGUGGAGUUCAAAACAAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGGCAGAUCUUCCUGGACCAGAACAUGCUGGCAGUGAUCGAUGAGCUGAUGCAAGCCUUGAACUUCAACUCAGAGACGGUGCCGCAAAAGUCCUCGUUGGAGGAACCAGAUUUUUACAAAACCAAAAUCAAGCUGUGUAUCCUUCUUCACGCCUUUCGGAUCAGAGCCGUGACUAUCGACCGGGUGAUGUCAUACCUGAAUGCUUCCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 340 hIL12AB_009UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAGCAGCUGGU 3′UTR)CAUCAGCUGGUUUAGCCUGGUCUUCCUGGCCAGCCCCCUGGUGGCCAUCUGGGAGCUGAAGAAAGACGUCUACGUAGUAGAGUUGGAUUGGUACCCAGACGCACCUGGAGAAAUGGUGGUUCUCACCUGCGACACGCCAGAAGAAGACGGUAUCACCUGGACGCUGGACCAGAGCAGCGAAGUACUGGGCAGUGGAAAAACGCUGACCAUACAAGUAAAAGAAUUUGGCGAUGCUGGCCAGUACACCUGCCACAAAGGAGGAGAAGUACUGAGCCACAGCCUGCUGCUGCUGCACAAGAAAGAAGAUGGCAUCUGGAGCACCGACAUUUUAAAAGACCAGAAGGAGCCCAAGAACAAAACCUUCCUUCGAUGUGAGGCGAAGAACUACAGUGGCCGCUUCACCUGCUGGUGGCUCACCACCAUCAGCACCGACCUCACCUUCUCGGUGAAGAGCAGCCGUGGUAGCUCAGACCCCCAAGGAGUCACCUGUGGGGCGGCCACGCUGUCGGCAGAAAGAGUUCGAGGCGACAACAAGGAAUAUGAAUACUCGGUGGAAUGUCAAGAAGAUUCGGCCUGCCCGGCGGCAGAAGAAAGUCUGCCCAUAGAAGUCAUGGUGGAUGCUGUUCACAAAUUAAAAUAUGAAAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAGCCAGACCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCCGGCAGGUGGAAGUUUCCUGGGAGUACCCAGAUACGUGGAGCACGCCGCACAGCUACUUCAGCCUCACCUUCUGUGUACAAGUACAAGGCAAGAGCAAGAGAGAGAAGAAAGAUCGUGUCUUCACCGACAAAACCUCGGCGACGGUCAUCUGCAGGAAGAAUGCAAGCAUCUCGGUUCGAGCCCAGGACCGCUACUACAGCAGCAGCUGGAGUGAGUGGGCCUCGGUGCCCUGCAGUGGUGGCGGCGGCGGCGGCAGCAGAAACCUUCCUGUGGCCACGCCGGACCCUGGCAUGUUUCCGUGCCUGCACCACAGCCAAAAUUUAUUACGAGCUGUUAGCAACAUGCUGCAGAAAGCACGGCAAACUUUAGAAUUCUACCCCUGCACCUCAGAAGAAAUAGACCAUGAAGAUAUCACCAAAGAUAAAACCAGCACUGUAGAGGCCUGCCUGCCCCUGGAGCUCACCAAGAACGAGAGCUGCCUCAAUAGCAGAGAGACCAGCUUCAUCACCAAUGGCAGCUGCCUGGCCAGCAGGAAAACCAGCUUCAUGAUGGCGCUCUGCCUGAGCAGCAUCUAUGAAGAUCUGAAGAUGUACCAAGUAGAAUUUAAAACCAUGAAUGCCAAGCUGCUCAUGGACCCCAAGCGGCAGAUAUUCCUCGACCAAAACAUGCUGGCUGUCAUUGAUGAGCUCAUGCAAGCAUUAAACUUCAACUCAGAGACGGUGCCCCAGAAGAGCAGCCUGGAGGAGCCAGAUUUCUACAAAACCAAGAUCAAGCUCUGCAUCUUAUUACAUGCCUUCCGCAUCCGGGCGGUCACCAUUGACCGUGUCAUGUCCUACUUAAAUGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 341 hIL12AB_010UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAGCAGCUUGU 3′UTR)CAUCUCCUGGUUUUCUCUUGUCUUCCUCGCUUCUCCUCUUGUGGCCAUCUGGGAGCUGAAGAAAGACGUCUACGUAGUAGAGUUGGAUUGGUACCCGGACGCUCCUGGAGAAAUGGUGGUUCUCACCUGCGACACUCCUGAAGAAGACGGUAUCACCUGGACGCUGGACCAAAGCAGCGAAGUUUUAGGCUCUGGAAAAACGCUGACCAUACAAGUAAAAGAAUUUGGCGACGCUGGCCAGUACACGUGCCACAAAGGAGGAGAAGUUUUAAGCCACAGUUUACUUCUUCUUCACAAGAAAGAAGAUGGCAUCUGGAGUACAGAUAUUUUAAAAGACCAGAAGGAGCCUAAGAACAAAACCUUCCUCCGCUGUGAAGCUAAGAACUACAGUGGUCGUUUCACCUGCUGGUGGCUCACCACCAUCUCCACUGACCUCACCUUCUCUGUAAAAUCAAGCCGUGGUUCUUCUGACCCCCAAGGAGUCACCUGUGGGGCUGCCACGCUCAGCGCUGAAAGAGUUCGAGGCGACAACAAGGAAUAUGAAUAUUCUGUGGAAUGUCAAGAAGAUUCUGCCUGCCCGGCGGCAGAAGAAAGUCUUCCCAUAGAAGUCAUGGUGGACGCUGUUCACAAAUUAAAAUAUGAAAACUACACCAGCAGCUUCUUCAUUCGUGACAUCAUCAAACCAGACCCUCCUAAGAACCUUCAGUUAAAACCGCUGAAGAACAGCCGGCAGGUGGAAGUUUCCUGGGAGUACCCAGAUACGUGGAGUACGCCGCACUCCUACUUCAGUUUAACCUUCUGUGUACAAGUACAAGGAAAAUCAAAAAGAGAGAAGAAAGAUCGUGUCUUCACUGACAAAACAUCUGCCACGGUCAUCUGCCGUAAGAACGCUUCCAUCUCGGUUCGAGCCCAGGACCGCUACUACAGCAGCAGCUGGAGUGAGUGGGCAUCUGUUCCCUGCAGUGGUGGCGGCGGCGGCGGCAGCCGCAACCUUCCUGUGGCCACGCCGGACCCUGGCAUGUUCCCGUGCCUUCACCACUCGCAAAAUCUUCUUCGUGCUGUUUCUAACAUGCUGCAGAAGGCGCGGCAAACUUUAGAAUUCUACCCGUGCACUUCUGAAGAAAUAGACCAUGAAGAUAUCACCAAAGAUAAAACCAGCACGGUGGAGGCCUGCCUUCCUUUAGAACUUACUAAGAACGAAAGUUGCCUUAACAGCCGUGAGACCAGCUUCAUCACCAAUGGCAGCUGCCUUGCUAGCAGGAAGACCAGCUUCAUGAUGGCGCUGUGCCUUUCUUCCAUCUAUGAAGAUCUUAAGAUGUACCAAGUAGAAUUUAAAACCAUGAAUGCCAAAUUAUUAAUGGACCCCAAGCGGCAGAUAUUCCUCGACCAAAACAUGCUGGCUGUCAUUGAUGAGCUCAUGCAAGCAUUAAACUUCAACUCAGAAACUGUUCCCCAGAAGUCAUCUUUAGAAGAACCAGAUUUCUACAAAACAAAAAUAAAACUCUGCAUUCUUCUUCAUGCCUUCCGCAUCCGUGCUGUCACCAUUGACCGUGUCAUGUCCUACUUAAAUGCUUCUUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 342 hIL12AB_011UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAGCAGCUGGU 3′UTR)GAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCAGCCCCCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGUUGGAUUGGUACCCGGACGCGCCGGGGGAGAUGGUGGUGCUGACGUGCGACACGCCGGAGGAGGACGGGAUCACGUGGACGCUGGACCAGAGCAGCGAGGUGCUGGGGAGCGGGAAGACGCUGACGAUCCAGGUGAAGGAGUUCGGGGACGCGGGGCAGUACACGUGCCACAAGGGGGGGGAGGUGCUGAGCCACAGCCUGCUGCUGCUGCACAAGAAGGAGGACGGGAUCUGGAGCACAGAUAUCCUGAAGGACCAGAAGGAGCCGAAGAACAAGACGUUCCUGAGGUGCGAGGCGAAGAACUACAGCGGGAGGUUCACGUGCUGGUGGCUGACGACGAUCAGCACGGACCUGACGUUCAGCGUGAAGAGCAGCAGGGGGAGCAGCGACCCGCAGGGGGUGACGUGCGGGGCGGCGACGCUGAGCGCGGAGAGGGUGAGGGGUGACAACAAGGAGUACGAGUACAGCGUGGAGUGCCAGGAAGAUAGCGCGUGCCCGGCGGCGGAGGAGAGCCUGCCGAUCGAGGUGAUGGUGGACGCGGUGCACAAGCUGAAGUACGAGAACUACACGAGCAGCUUCUUCAUCAGAGAUAUCAUCAAGCCGGACCCGCCGAAGAACCUGCAGCUGAAGCCGCUGAAGAACAGCAGGCAGGUGGAGGUGAGCUGGGAGUACCCAGAUACGUGGAGCACGCCGCACAGCUACUUCAGCCUGACGUUCUGCGUGCAGGUGCAGGGGAAGAGCAAGAGGGAGAAGAAAGAUAGGGUGUUCACAGAUAAGACGAGCGCGACGGUGAUCUGCAGGAAGAACGCGAGCAUCAGCGUGAGGGCGCAAGAUAGGUACUACAGCAGCAGCUGGAGCGAGUGGGCGAGCGUGCCGUGCAGCGGGGGGGGGGGGGGGGGGAGCAGGAACCUGCCGGUGGCGACGCCGGACCCGGGGAUGUUCCCGUGCCUGCACCACAGCCAGAACCUGCUGAGGGCGGUGAGCAACAUGCUGCAGAAGGCGAGGCAGACGCUGGAGUUCUACCCGUGCACGAGCGAGGAGAUCGACCACGAAGAUAUCACGAAAGAUAAGACGAGCACGGUGGAGGCGUGCCUGCCGCUGGAGCUGACGAAGAACGAGAGCUGCCUGAACAGCAGGGAGACGAGCUUCAUCACGAACGGGAGCUGCCUGGCGAGCAGGAAGACGAGCUUCAUGAUGGCGCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACGAUGAACGCGAAGCUGCUGAUGGACCCGAAGAGGCAGAUCUUCCUGGACCAGAACAUGCUGGCGGUGAUCGACGAGCUGAUGCAGGCGCUGAACUUCAACAGCGAGACGGUGCCGCAGAAGAGCAGCCUGGAGGAGCCAGAUUUCUACAAGACGAAGAUCAAGCUGUGCAUCCUGCUGCACGCGUUCAGGAUCAGGGCGGUGACGAUCGACAGGGUGAUGAGCUACCUGAACGCGAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 343 hIL12AB_012UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCAUCAGCAGCUGGU 3′UTR)GAUCAGCUGGUUCAGCCUCGUGUUUCUGGCCAGCCCCCUGGUGGCCAUUUGGGAACUCAAGAAGGACGUGUACGUUGUGGAACUCGACUGGUACCCUGACGCCCCAGGCGAAAUGGUGGUCUUAACCUGCGACACCCCUGAGGAGGACGGAAUCACCUGGACCUUGGACCAGAGCUCCGAGGUCCUCGGCAGUGGCAAGACCCUGACCAUACAGGUGAAAGAAUUUGGAGACGCAGGGCAAUACACAUGUCACAAGGGCGGGGAGGUUCUUUCUCACUCCCUUCUGCUUCUACAUAAAAAGGAAGACGGAAUUUGGUCUACCGACAUCCUCAAGGACCAAAAGGAGCCUAAGAAUAAAACCUUCUUACGCUGUGAAGCUAAAAACUACAGCGGCAGAUUCACUUGCUGGUGGCUCACCACCAUUUCUACCGACCUGACCUUCUCGGUGAAGUCUUCAAGGGGCUCUAGUGAUCCACAGGGAGUGACAUGCGGGGCCGCCACACUGAGCGCUGAACGGGUGAGGGGCGAUAACAAGGAGUAUGAAUACUCUGUCGAGUGUCAGGAGGAUUCAGCUUGUCCCGCAGCUGAAGAGUCACUCCCCAUAGAGGUUAUGGUCGAUGCUGUGCAUAAACUGAAGUACGAAAACUACACCAGCAGCUUCUUCAUUAGAGAUAUUAUAAAACCUGAGCCCCCCAAGAACCUGCAACUUAAACCCCUGAAAAACUCUCGGCAGGUCGAAGUUAGCUGGGAGUACCCUGAUACUUGGUCCACCCCCCACUCGUACUUCUCACUGACUUUCUGUGUGCAGGUGCAGGGCAAGAGCAAGAGAGAGAAAAAAGAUCGUGUAUUCACAGAUAAGACCUCUGCCACCGUGAUCUGCAGAAAAAACGCUUCCAUCAGUGUCAGAGCCCAAGACCGGUACUAUAGUAGUAGCUGGAGCGAGUGGGCAAGUGUCCCCUGCUCUGGCGGCGGAGGGGGCGGCUCUCGAAACCUCCCCGUCGCUACCCCUGAUCCAGGAAUGUUCCCUUGCCUGCAUCACUCACAGAAUCUGCUGAGAGCGGUCAGCAACAUGCUGCAGAAAGCUAGGCAAACACUGGAGUUUUAUCCUUGUACCUCAGAGGAGAUCGACCACGAGGAUAUUACCAAAGAUAAGACCAGCACGGUGGAGGCCUGCUUGCCCCUGGAACUGACAAAGAAUGAAUCCUGCCUUAAUAGCCGUGAGACCUCUUUUAUAACAAACGGAUCCUGCCUGGCCAGCAGGAAGACCUCCUUCAUGAUGGCCCUCUGCCUGUCCUCAAUCUACGAAGACCUGAAGAUGUACCAGGUGGAAUUUAAAACUAUGAACGCCAAGCUGUUGAUGGACCCCAAGCGGCAGAUCUUUCUGGAUCAAAAUAUGCUGGCUGUGAUCGACGAACUGAUGCAGGCCCUCAACUUUAACAGCGAGACCGUGCCACAAAAGAGCAGUCUUGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUCCUUCAUGCCUUCAGGAUAAGAGCUGUCACCAUCGACAGAGUCAUGAGUUACCUGAAUGCAUCCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 344 hIL12AB_013UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAGCAGCUGGU 3′UTR)CAUCUCCUGGUUCAGUCUUGUCUUCCUGGCCUCGCCGCUGGUGGCCAUCUGGGAGCUGAAGAAAGACGUUUACGUAGUAGAGUUGGAUUGGUACCCAGACGCACCUGGAGAAAUGGUGGUCCUCACCUGUGACACGCCAGAAGAAGACGGUAUCACCUGGACGCUGGACCAGAGCAGUGAAGUUCUUGGAAGUGGAAAAACGCUGACCAUACAAGUAAAAGAAUUUGGAGAUGCUGGCCAGUACACCUGCCACAAAGGAGGAGAAGUUCUCAGCCACAGUUUAUUAUUACUUCACAAGAAAGAAGAUGGCAUCUGGUCCACAGAUAUUUUAAAAGACCAGAAGGAGCCCAAAAAUAAAACAUUUCUUCGAUGUGAGGCCAAGAACUACAGUGGUCGUUUCACCUGCUGGUGGCUGACCACCAUCUCCACAGACCUCACCUUCAGUGUAAAAAGCAGCCGUGGUUCUUCUGACCCCCAAGGAGUCACCUGUGGGGCUGCCACGCUCUCUGCAGAAAGAGUUCGAGGUGACAACAAAGAAUAUGAGUACUCGGUGGAAUGUCAAGAAGAUUCGGCCUGCCCAGCUGCUGAGGAGAGUCUUCCCAUAGAAGUCAUGGUGGAUGCUGUUCACAAAUUAAAAUAUGAAAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAACCUGACCCGCCCAAGAACUUACAGCUGAAGCCGCUGAAAAACAGCCGGCAGGUAGAAGUUUCCUGGGAGUACCCAGAUACCUGGUCCACGCCGCACUCCUACUUCUCCCUCACCUUCUGUGUACAAGUACAAGGCAAGAGCAAGAGAGAGAAGAAAGAUCGUGUCUUCACAGAUAAAACAUCAGCCACGGUCAUCUGCAGGAAAAAUGCCAGCAUCUCGGUGCGGGCCCAGGACCGCUACUACAGCAGCAGCUGGAGUGAGUGGGCAUCUGUGCCCUGCAGUGGUGGUGGGGGUGGUGGCAGCAGAAACCUUCCUGUGGCCACUCCAGACCCUGGCAUGUUCCCGUGCCUUCACCACUCCCAAAAUUUACUUCGAGCUGUUUCUAACAUGCUGCAGAAAGCACGGCAAACUUUAGAAUUCUACCCGUGCACUUCUGAAGAAAUUGACCAUGAAGAUAUCACAAAAGAUAAAACCAGCACAGUGGAGGCCUGUCUUCCUUUAGAGCUGACCAAAAAUGAAUCCUGCCUCAACAGCAGAGAGACCAGCUUCAUCACCAAUGGCAGCUGCCUGGCCUCCAGGAAAACCAGCUUCAUGAUGGCGCUCUGCCUCAGCUCCAUCUAUGAAGAUUUGAAGAUGUACCAAGUAGAAUUUAAAACCAUGAAUGCCAAAUUAUUAAUGGACCCCAAGAGGCAGAUAUUUUUAGAUCAAAACAUGCUGGCAGUUAUUGAUGAGCUCAUGCAAGCAUUAAACUUCAACAGUGAGACGGUACCUCAAAAAAGCAGCCUUGAAGAGCCAGAUUUCUACAAAACCAAGAUCAAACUCUGCAUUUUACUUCAUGCCUUCCGCAUCCGGGCGGUCACCAUUGACCGUGUCAUGUCCUACUUAAAUGCCUCGUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 345 hIL12AB_014UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAGCAGCUUGU 3′UTR)GAUUUCUUGGUUCUCUCUUGUGUUCCUUGCUUCUCCUCUUGUGGCUAUUUGGGAGUUAAAAAAGGACGUGUACGUGGUGGAGCUUGACUGGUACCCUGACGCACCUGGCGAGAUGGUGGUGCUUACUUGUGACACUCCUGAGGAGGACGGCAUUACUUGGACGCUUGACCAGUCUUCUGAGGUGCUUGGCUCUGGCAAAACACUUACUAUUCAGGUGAAGGAGUUCGGGGAUGCUGGCCAGUACACUUGCCACAAGGGCGGCGAGGUGCUUUCUCACUCUCUUCUUCUUCUUCACAAGAAGGAGGACGGCAUUUGGUCUACUGACAUUUUAAAAGACCAGAAGGAGCCCAAGAACAAAACAUUCCUUCGUUGCGAGGCCAAGAACUACUCUGGCCGUUUCACUUGCUGGUGGCUUACUACUAUUUCUACUGACCUUACUUUCUCUGUGAAGUCUUCUCGUGGCUCUUCUGACCCUCAGGGCGUGACUUGUGGGGCUGCUACUCUUUCUGCUGAGCGUGUGCGUGGUGACAACAAGGAGUACGAGUACUCUGUGGAGUGCCAGGAAGAUUCUGCUUGCCCUGCUGCUGAGGAGUCUCUUCCUAUUGAGGUGAUGGUGGAUGCUGUGCACAAGUUAAAAUACGAGAACUACACUUCUUCUUUCUUCAUUCGUGACAUUAUUAAGCCUGACCCUCCCAAGAACCUUCAGUUAAAACCUUUAAAAAACUCUCGUCAGGUGGAGGUGUCUUGGGAGUACCCUGACACUUGGUCUACUCCUCACUCUUACUUCUCUCUUACUUUCUGCGUGCAGGUGCAGGGCAAGUCUAAGCGUGAGAAGAAGGACCGUGUGUUCACUGACAAAACAUCUGCUACUGUGAUUUGCAGGAAGAAUGCAUCUAUUUCUGUGCGUGCUCAGGACCGUUACUACUCUUCUUCUUGGUCUGAGUGGGCUUCUGUGCCUUGCUCUGGCGGCGGCGGCGGCGGCUCCAGAAAUCUUCCUGUGGCUACUCCUGACCCUGGCAUGUUCCCUUGCCUUCACCACUCUCAGAACCUUCUUCGUGCUGUGAGCAACAUGCUUCAGAAGGCUCGUCAAACUCUUGAGUUCUACCCUUGCACUUCUGAGGAGAUUGAGCACGAAGAUAUCACCAAAGAUAAAACAUCUACUGUGGAGGCUUGCCUUCGUCUUGAGCUUACCAAGAAUGAAUCUUGCUUAAAUUCUCGUGAGACGUCUUUGAUCACCAACGGCUCUUGCCUUGCCUCGCGCAAAACAUCUUUCAUGAUGGCUCUUUGCCUUUCUUCUAUUUACGAAGAUUUAAAAAUGUACCAGGUGGAGUUCAAAACAAUGAAUGCAAAGCUUCUUAUGGACCCCAAGCGUCAGAUUUUCCUUGACCAGAACAUGCUUGCUGUGAUUGACGAGCUUAUGCAGGCUUUAAAUUUCAACUCUGAGACGGUGCCUCAGAAGUCUUCUCUUGAGGAGCCUGACUUCUACAAGACCAAGAUUAAGCUUUGCAUUCUUCUUCAUGCUUUCCGUAUUCGUGCUGUGACUAUUGACCGUGUGAUGUCUUACUUAAAUGCUUCUUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 346 hIL12AB_015UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGUCACCAGCAGCUGGU 3′UTR)GAUCAGCUGGUUUAGCCUGGUGUUUCUGGCCAGCCCCCUGGUGGCCAUCUGGGAACUGAAGAAAGACGUGUACGUGGUAGAACUGGAUUGGUAUCCGGACGCUCCCGGCGAAAUGGUGGUGCUGACCUGUGACACCCCCGAAGAAGACGGAAUCACCUGGACCCUGGACCAGAGCAGCGAGGUGCUGGGCAGCGGCAAAACCCUGACCAUCCAAGUGAAAGAGUUUGGCGAUGCCGGCCAGUACACCUGUCACAAAGGCGGCGAGGUGCUAAGCCAUUCGCUGCUGCUGCUGCACAAAAAGGAAGAUGGCAUCUGGAGCACCGAUAUCCUGAAGGACCAGAAAGAACCCAAAAAUAAGACCUUUCUAAGAUGCGAGGCCAAGAAUUAUAGCGGCCGUUUCACCUGCUGGUGGCUGACGACCAUCAGCACCGAUCUGACCUUCAGCGUGAAAAGCAGCAGAGGCAGCAGCGACCCCCAAGGCGUGACGUGCGGCGCCGCCACCCUGAGCGCCGAGAGAGUGAGAGGCGACAACAAGGAGUAUGAGUACAGCGUGGAGUGCCAGGAAGAUAGCGCCUGCCCCGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGAUGCCGUGCACAAGCUGAAGUAUGAAAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAACCCGACCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAAUAGCCGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGAGCACCCCCCAUAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGCAAGAGCAAGAGAGAAAAGAAAGAUAGAGUGUUCACAGAUAAGACCAGCGCCACGGUGAUCUGCAGAAAAAAUGCCAGCAUCAGCGUGAGAGCCCAAGAUAGAUACUAUAGCAGCAGCUGGAGCGAAUGGGCCAGCGUGCCCUGCAGCGGCGGCGGCGGCGGCGGCAGCAGAAACCUGCCCGUGGCCACCCCCGACCCCGGCAUGUUCCCCUGCCUGCACCACAGCCAAAACCUGCUGAGAGCCGUGAGCAACAUGCUGCAGAAGGCCCGGCAGACCCUGGAAUUUUACCCCUGCACCAGCGAAGAGAUCGAUCAUGAAGAUAUCACCAAAGAUAAAACCAGCACCGUGGAGGCCUGUCUGCCCCUGGAACUGACCAAGAAUGAGAGCUGCCUAAAUAGCAGAGAGACCAGCUUCAUAACCAAUGGCAGCUGCCUGGCCAGCAGAAAGACCAGCUUUAUGAUGGCCCUGUGCCUGAGCAGCAUCUAUGAAGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAAUGCCAAGCUGCUGAUGGAUCCCAAGCGGCAGAUCUUUCUGGAUCAAAACAUGCUGGCCGUGAUCGAUGAGCUGAUGCAGGCCCUGAAUUUCAACAGCGAGACCGUGCCCCAAAAAAGCAGCCUGGAAGAACCGGAUUUUUAUAAAACCAAAAUCAAGCUGUGCAUACUGCUGCAUGCCUUCAGAAUCAGAGCCGUGACCAUCGAUAGAGUGAUGAGCUAUCUGAAUGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 347 hIL12AB_016UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAGCAGCUGGU 3′UTR)CAUCAGCUGGUUCAGCCUGGUCUUCCUGGCCAGCCCCCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUAUACGUAGUGGAGUUGGAUUGGUACCCAGACGCUCCUGGGGAGAUGGUGGUGCUGACCUGUGACACCCCAGAAGAGGACGGUAUCACCUGGACCCUGGACCAGAGCUCAGAAGUGCUGGGCAGUGGAAAAACCCUGACCAUCCAGGUGAAGGAGUUUGGAGAUGCUGGCCAGUACACCUGCCACAAGGGUGGUGAAGUGCUGAGCCACAGCCUGCUGCUGCUGCACAAGAAGGAGGAUGGCAUCUGGAGCACAGAUAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACCUUCCUUCGCUGUGAAGCCAAGAACUACAGUGGCCGCUUCACCUGCUGGUGGCUGACCACCAUCAGCACAGACCUCACCUUCUCGGUGAAGAGCAGCAGAGGCAGCUCAGACCCCCAGGGUGUCACCUGUGGGGCGGCCACGCUGUCGGCGGAGAGAGUUCGAGGUGACAACAAGGAGUAUGAAUACUCGGUGGAGUGCCAGGAAGAUUCGGCGUGCCCGGCGGCAGAAGAGAGCCUGCCCAUAGAAGUGAUGGUGGAUGCUGUGCACAAGCUGAAGUAUGAAAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAGCCAGACCCGCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCCGGCAGGUGGAGGUUUCCUGGGAGUACCCAGAUACGUGGAGCACCCCCCACAGCUACUUCAGCCUGACCUUCUGUGUCCAGGUGCAGGGCAAGAGCAAGAGAGAGAAGAAAGAUAGAGUCUUCACAGAUAAGACCUCGGCCACGGUCAUCUGCAGAAAGAAUGCCUCCAUCUCGGUUCGAGCCCAAGAUAGAUACUACAGCAGCAGCUGGUCAGAAUGGGCCUCGGUGCCCUGCAGUGGUGGCGGCGGCGGCGGCAGCAGAAACCUGCCUGUUGCCACCCCAGACCCUGGGAUGUUCCCCUGCCUGCACCACAGCCAGAACUUAUUACGAGCUGUUUCUAACAUGCUGCAGAAGGCCCGGCAGACCCUGGAGUUCUACCCCUGCACCUCAGAAGAGAUUGACCAUGAAGAUAUCACCAAAGAUAAGACCAGCACUGUAGAGGCCUGCCUGCCCCUGGAGCUGACCAAGAAUGAAAGCUGCCUGAACAGCAGAGAGACCAGCUUCAUCACCAAUGGAAGCUGCCUGGCCAGCAGAAAGACCAGCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUAUGAAGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAAUGCAAAGCUGCUGAUGGACCCCAAGCGGCAGAUAUUUUUGGACCAGAACAUGCUGGCUGUCAUUGAUGAGCUGAUGCAGGCCCUGAACUUCAACUCAGAAACUGUACCCCAGAAGAGCAGCCUGGAGGAGCCAGAUUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUUCAUGCUUUCAGAAUCAGAGCUGUCACCAUUGACCGCGUGAUGAGCUACUUAAAUGCCUCGUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 348 hIL12AB_017UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAGCAGCUGGU 3′UTR)AAUCAGCUGGUUUUCCCUCGUCUUUCUGGCAUCACCCCUGGUGGCUAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGCUGGAUUGGUACCCUGACGCCCCGGGGGAAAUGGUGGUGUUAACCUGCGACACGCCUGAGGAGGACGGCAUCACCUGGACGCUGGACCAGAGCAGCGAGGUGCUUGGGUCUGGUAAAACUCUGACUAUUCAGGUGAAAGAGUUCGGGGAUGCCGGCCAAUAUACUUGCCACAAGGGUGGCGAGGUGCUUUCUCAUUCUCUGCUCCUGCUGCACAAGAAAGAAGAUGGCAUUUGGUCUACUGAUAUUCUGAAAGACCAGAAGGAGCCCAAGAACAAGACCUUUCUGAGAUGCGAGGCUAAAAACUACAGCGGAAGAUUUACCUGCUGGUGGCUGACCACAAUCUCAACCGACCUGACAUUUUCAGUGAAGUCCAGCAGAGGGAGCUCCGACCCUCAGGGCGUGACCUGCGGAGCCGCCACUCUGUCCGCAGAAAGAGUGAGAGGUGAUAAUAAGGAGUACGAGUAUUCAGUCGAGUGCCAAGAAGAUUCUGCCUGCCCAGCCGCCGAGGAGAGCCUGCCAAUCGAGGUGAUGGUAGAUGCGGUACACAAGCUGAAGUAUGAGAACUACACAUCCUCCUUCUUCAUAAGAGAUAUUAUCAAGCCUGACCCACCUAAAAAUCUGCAACUCAAGCCUUUGAAAAAUUCACGGCAGGUGGAGGUGAGCUGGGAGUACCCUGAUACUUGGAGCACCCCCCAUAGCUACUUUUCGCUGACAUUCUGCGUCCAGGUGCAGGGCAAGUCAAAGAGAGAGAAGAAGGAUCGCGUGUUCACUGAUAAAACAAGCGCCACAGUGAUCUGCAGAAAAAACGCUAGCAUUAGCGUCAGAGCACAGGACCGGUAUUACUCCAGCUCCUGGAGCGAAUGGGCAUCUGUGCCCUGCAGCGGUGGGGGCGGAGGCGGAUCCAGAAACCUCCCCGUUGCCACACCUGAUCCUGGAAUGUUCCCCUGUCUGCACCACAGCCAGAACCUGCUGAGAGCAGUGUCUAACAUGCUCCAGAAGGCCAGGCAGACCCUGGAGUUUUACCCCUGCACCAGCGAGGAAAUCGAUCACGAAGAUAUCACCAAAGAUAAAACCUCCACCGUGGAGGCCUGCCUGCCCCUGGAACUGACCAAAAACGAGAGCUGCCUGAAUAGCAGGGAGACCUCCUUCAUCACCAACGGCUCAUGCCUUGCCAGCCGGAAAACUAGCUUCAUGAUGGCCCUGUGCCUGUCUUCGAUCUAUGAGGACCUGAAAAUGUACCAGGUCGAAUUUAAGACGAUGAACGCAAAGCUGCUGAUGGACCCCAAGCGGCAGAUCUUUCUGGACCAGAACAUGCUGGCAGUCAUAGAUGAGUUGAUGCAGGCAUUAAACUUCAACAGCGAGACCGUGCCUCAGAAGUCCAGCCUCGAGGAGCCAGAUUUUUAUAAGACCAAGAUCAAACUAUGCAUCCUGCUGCAUGCUUUCAGGAUUAGAGCCGUCACCAUCGAUCGAGUCAUGUCUUACCUGAAUGCUAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 349 hIL12AB_018UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGUCACCAACAGUUAGU 3′UTR)AAUCUCCUGGUUUUCUCUGGUGUUUCUGGCCAGCCCCCUCGUGGCCAUCUGGGAGCUUAAAAAGGACGUUUACGUGGUGGAGUUGGAUUGGUAUCCCGACGCUCCAGGCGAAAUGGUCGUGCUGACCUGCGAUACCCCUGAAGAAGACGGUAUCACCUGGACGCUGGACCAGUCUUCCGAGGUGCUUGGAUCUGGCAAAACACUGACAAUACAAGUUAAGGAGUUCGGGGACGCAGGGCAGUACACCUGCCACAAAGGCGGCGAGGUCCUGAGUCACUCCCUGUUACUGCUCCACAAGAAAGAGGACGGCAUUUGGUCCACCGACAUUCUGAAGGACCAGAAGGAGCCUAAGAAUAAAACUUUCCUGAGAUGCGAGGCAAAAAACUAUAGCGGCCGCUUUACUUGCUGGUGGCUUACAACAAUCUCUACCGAUUUAACUUUCUCCGUGAAGUCUAGCAGAGGAUCCUCUGACCCGCAAGGAGUGACUUGCGGAGCCGCCACCUUGAGCGCCGAAAGAGUCCGUGGCGAUAACAAAGAAUACGAGUACUCCGUGGAGUGCCAGGAAGAUUCCGCCUGCCCAGCUGCCGAGGAGUCCCUGCCCAUUGAAGUGAUGGUGGAUGCCGUCCACAAGCUGAAGUACGAAAACUAUACCAGCAGCUUCUUCAUCCGGGAUAUCAUUAAGCCCGACCCUCCUAAAAACCUGCAACUUAAGCCCCUAAAGAAUAGUCGGCAGGUUGAGGUCAGCUGGGAAUAUCCUGACACAUGGAGCACCCCCCACUCUUAUUUCUCCCUGACCUUCUGCGUGCAGGUGCAGGGCAAGAGUAAACGGGAGAAAAAAGAUAGGGUCUUUACCGAUAAAACCAGCGCUACGGUUAUCUGUCGGAAGAACGCUUCCAUCUCCGUCCGCGCUCAGGAUCGUUACUACUCGUCCUCAUGGAGCGAGUGGGCCAGCGUGCCCUGCAGCGGCGGCGGUGGAGGCGGAUCCAGAAAUCUGCCUGUUGCCACACCAGACCCUGGCAUGUUCCCCUGUCUGCAUCAUAGCCAGAACCUGCUCAGAGCCGUGAGCAACAUGCUCCAGAAGGCCAGGCAAACUUUGGAGUUCUACCCGUGUACAUCUGAGGAAAUCGAUCACGAAGAUAUAACCAAAGAUAAAACCUCUACAGUAGAGGCUUGUUUGCCCCUGGAGUUGACCAAAAACGAGAGUUGCCUGAACAGUCGCGAGACGAGCUUCAUUACUAACGGCAGCUGUCUCGCCUCCAGAAAAACAUCCUUCAUGAUGGCCCUGUGUCUUUCCAGCAUAUACGAAGACCUGAAAAUGUACCAGGUCGAGUUCAAAACAAUGAACGCCAAGCUGCUUAUGGACCCCAAGCGGCAGAUCUUCCUCGACCAAAACAUGCUCGCUGUGAUCGAUGAGCUGAUGCAGGCUCUCAACUUCAAUUCCGAAACAGUGCCACAGAAGUCCAGUCUGGAAGAACCCGACUUCUACAAGACCAAGAUUAAGCUGUGUAUUUUGCUGCAUGCGUUUAGAAUCAGAGCCGUGACCAUUGAUCGGGUGAUGAGCUACCUGAACGCCUCGUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 350 hIL12AB_019UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAGCAGCUUGU 3′UTR)CAUCUCCUGGUUUUCUCUUGUCUUCCUGGCCUCGCCGCUGGUGGCCAUCUGGGAGCUGAAGAAAGACGUUUACGUAGUAGAGUUGGAUUGGUACCCAGACGCACCUGGAGAAAUGGUGGUUCUCACCUGUGACACUCCUGAAGAAGACGGUAUCACCUGGACGCUGGACCAAAGCUCAGAAGUUCUUGGCAGUGGAAAAACGCUGACCAUACAAGUAAAAGAAUUUGGGGAUGCUGGCCAGUACACGUGCCACAAAGGAGGAGAAGUUCUCAGCCACAGUUUACUUCUUCUUCACAAGAAAGAAGAUGGCAUCUGGUCCACAGAUAUUUUAAAAGACCAGAAGGAGCCCAAGAACAAAACCUUCCUCCGCUGUGAGGCCAAGAACUACAGUGGUCGUUUCACCUGCUGGUGGCUCACCACCAUCUCCACUGACCUCACCUUCUCUGUAAAAAGCAGCCGUGGUUCUUCUGACCCCCAAGGAGUCACCUGUGGGGCUGCCACGCUCUCGGCAGAAAGAGUUCGAGGUGACAACAAGGAAUAUGAAUAUUCUGUGGAAUGUCAAGAAGAUUCUGCCUGCCCGGCGGCAGAAGAAAGUCUUCCCAUAGAAGUCAUGGUGGAUGCUGUUCACAAAUUAAAAUAUGAAAACUACACCAGCAGCUUCUUCAUUCGUGACAUCAUCAAACCAGACCCGCCCAAGAACCUUCAGUUAAAACCUUUAAAAAACAGCCGGCAGGUAGAAGUUUCCUGGGAGUACCCAGAUACGUGGUCCACGCCGCACUCCUACUUCAGUUUAACCUUCUGUGUACAAGUACAAGGAAAAUCAAAAAGAGAGAAGAAAGAUCGUGUCUUCACUGACAAAACAUCUGCCACGGUCAUCUGCAGGAAGAAUGCCUCCAUCUCGGUUCGAGCCCAGGACCGCUACUACAGCAGCAGCUGGAGUGAGUGGGCAUCUGUUCCCUGCAGUGGUGGCGGCGGCGGCGGCAGCCGCAACCUUCCUGUGGCCACGCCGGACCCUGGCAUGUUCCCGUGCCUUCACCACUCCCAAAAUCUUCUUCGUGCUGUUUCUAACAUGCUGCAGAAGGCGCGCCAAACUUUAGAAUUCUACCCGUGCACUUCUGAAGAAAUAGACCAUGAAGAUAUCACCAAAGAUAAAACCAGCACGGUGGAGGCCUGCCUUCCUUUAGAGCUGAGCAAGAAUGAAUCCUGCCUCAACAGCAGAGAGACCAGCUUCAUCACCAAUGGCAGCUGCCUGGCCUCGCGCAAGACCAGCUUCAUGAUGGCGCUGUGCCUUUCUUCCAUCUAUGAAGAUUUAAAGAUGUACCAAGUAGAAUUUAAAACCAUGAAUGCCAAAUUAUUAAUGGACCCCAAACGGCAGAUAUUUUUGGAUCAAAACAUGCUGGCUGUCAUUGAUGAGCUCAUGCAAGCAUUAAACUUCAACUCAGAAACUGUUCCCCAGAAGUCAUCUUUAGAAGAGCCAGAUUUCUACAAAACAAAAAUAAAACUCUGCAUUCUUCUUCAUGCCUUCCGCAUCCGUGCUGUCACCAUUGACCGUGUCAUGUCCUACUUAAAUGCUUCUUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 351 hIL12AB_020UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAGCAGCUGGU 3′UTR)GAUCAGCUGGUUCAGCCUGGUGUUCCUGGCUAGCCCUCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGUUGGAUUGGUACCCCGACGCUCCCGGCGAGAUGGUGGUGCUGACCUGCGACACCCCCGAGGAGGACGGGAUCACCUGGACCCUGGAUCAGUCAAGCGAGGUGCUGGGAAGCGGCAAGACCCUGACCAUCCAGGUGAAGGAGUUCGGCGACGCCGGCCAAUACACUUGCCACAAGGGAGGCGAGGUGCUGUCCCACUCCCUCCUGCUGCUGCACAAAAAGGAAGACGGCAUCUGGAGCACCGACAUCCUGAAAGACCAGAAGGAGCCUAAGAACAAAACAUUCCUCAGAUGCGAGGCCAAGAAUUACUCCGGGAGAUUCACCUGUUGGUGGCUGACCACCAUCAGCACAGACCUGACCUUCAGCGUGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGUGACCUGUGGCGCCGCCACCCUGAGCGCCGAAAGAGUGCGCGGCGACAACAAGGAGUACGAGUACUCCGUGGAAUGCCAGGAAGAUAGCGCCUGCCCCGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUCCACAAGCUGAAGUACGAGAACUACACCUCUAGCUUCUUCAUCAGAGAUAUCAUCAAGCCCGAUCCCCCCAAGAACCUGCAGCUGAAACCCCUGAAGAACAGCCGGCAGGUGGAGGUGAGCUGGGAGUAUCCCGACACCUGGUCCACCCCCCACAGCUAUUUUAGCCUGACCUUCUGCGUGCAAGUGCAGGGCAAGAGCAAGAGAGAGAAGAAGGACCGCGUGUUCACCGACAAAACCAGCGCCACCGUGAUCUGCAGAAAGAACGCCAGCAUCAGCGUGAGGGCCCAGGAUAGAUACUACAGUUCCAGCUGGAGCGAGUGGGCCAGCGUGCCCUGCAGCGGCGGCGGCGGGGGAGGCUCGAGAAACCUGCCCGUGGCUACCCCCGAUCCCGGAAUGUUCCCCUGCCUGCACCACAGCCAGAACCUGCUGAGGGCGGUGUCCAACAUGCUUCAGAAGGCCCGGCAGACCCUGGAGUUCUACCCCUGUACCUCUGAGGAGAUCGAUCAUGAAGAUAUCACAAAAGAUAAAACCAGCACCGUGGAGGCCUGCCUGCCCCUGGAGCUGACCAAGAACGAGAGCUGCCUGAACUCCCGCGAGACCAGCUUCAUCACGAACGGCAGCUGCCUGGCCAGCAGGAAGACCUCCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAAAUGUACCAGGUGGAGUUUAAGACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGGCAAAUCUUCCUGGACCAGAACAUGCUGGCAGUGAUCGACGAGCUCAUGCAGGCCCUGAACUUCAAUAGCGAGACGGUCCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUUUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUUAGAAUCCGUGCCGUGACCAUUGACAGAGUGAUGAGCUACCUGAAUGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 352 hIL12AB_021UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAGCAGCUGGU 3′UTR)GAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCAGCCCUCUGGUUGCCAUCUGGGAGCUGAAGAAAGACGUGUACGUCGUGGAACUGGACUGGUAUCCGGACGCCCCGGGCGAGAUGGUGGUGCUGACCUGUGACACCCCCGAGGAGGACGGCAUCACCUGGACGCUGGACCAAUCCUCCGAGGUGCUGGGAAGCGGCAAGACCCUGACCAUCCAGGUGAAGGAAUUCGGGGACGCCGGGCAGUACACCUGCCACAAGGGGGGCGAAGUGCUGUCCCACUCGCUGCUGCUCCUGCAUAAGAAGGAGGAUGGAAUCUGGUCCACCGACAUCCUCAAAGAUCAGAAGGAGCCCAAGAACAAGACGUUCCUGCGCUGUGAAGCCAAGAAUUAUUCGGGGCGAUUCACGUGCUGGUGGCUGACAACCAUCAGCACCGACCUGACGUUUAGCGUGAAGAGCAGCAGGGGGUCCAGCGACCCCCAGGGCGUGACGUGCGGCGCCGCCACCCUCUCCGCCGAGAGGGUGCGGGGGGACAAUAAGGAGUACGAGUACAGCGUGGAAUGCCAGGAGGACAGCGCCUGCCCCGCCGCGGAGGAAAGCCUCCCGAUAGAGGUGAUGGUGGACGCCGUGCACAAGCUCAAGUAUGAGAAUUACACCAGCAGCUUUUUCAUCCGGGACAUUAUCAAGCCCGACCCCCCGAAGAACCUCCAGCUGAAGCCCCUGAAGAACAGCCGGCAGGUGGAAGUCUCCUGGGAGUAUCCCGACACCUGGAGCACCCCGCACAGCUACUUCUCCCUGACCUUCUGUGUGCAGGUGCAGGGCAAGUCCAAGAGGGAAAAGAAGGACAGGGUUUUCACCGACAAGACCAGCGCGACCGUGAUCUGCCGGAAGAACGCCAGCAUAAGCGUCCGCGCCCAAGAUAGGUACUACAGCAGCUCCUGGAGCGAGUGGGCUAGCGUGCCCUGCAGCGGGGGCGGGGGUGGGGGCUCCAGGAACCUGCCAGUGGCGACCCCCGACCCCGGCAUGUUCCCCUGCCUCCAUCACAGCCAGAACCUGCUGAGGGCCGUCAGCAAUAUGCUGCAGAAGGCCAGGCAGACCCUGGAAUUCUACCCCUGCACGUCGGAGGAGAUCGAUCACGAGGAUAUCACAAAAGACAAGACUUCCACCGUGGAGGCCUGCCUGCCCCUGGAGCUCACCAAGAAUGAGUCCUGUCUGAACUCCCGGGAAACCAGCUUCAUCACCAACGGGUCCUGCCUGGCCAGCAGGAAGACCAGCUUUAUGAUGGCCCUGUGCCUGUCGAGCAUCUACGAGGACCUGAAGAUGUACCAGGUCGAGUUCAAGACAAUGAACGCCAAGCUGCUGAUGGACCCCAAGAGGCAAAUCUUCCUGGACCAGAAUAUGCUUGCCGUCAUCGACGAGCUCAUGCAGGCCCUGAACUUCAACUCCGAGACCGUGCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCGUUCAGGAUCCGGGCAGUCACCAUCGACCGUGUGAUGUCCUACCUGAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 353 hIL12AB_022UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCAUCAGCAGCUGGU 3′UTR)GAUCAGCUGGUUCAGCCUGGUGUUCCUCGCCUCUCCCCUGGUGGCCAUCUGGGAGCUCAAAAAGGACGUGUACGUGGUGGAGCUCGACUGGUACCCAGACGCCCCCGGGGAGAUGGUGGUGCUGACCUGCGACACCCCCGAAGAAGACGGCAUCACGUGGACCCUCGACCAGUCCAGCGAGGUGCUGGGGAGCGGGAAGACUCUGACCAUCCAGGUCAAGGAGUUCGGGGACGCCGGGCAGUACACGUGCCACAAGGGCGGCGAAGUCUUAAGCCACAGCCUGCUCCUGCUGCACAAGAAGGAGGACGGGAUCUGGUCCACAGACAUACUGAAGGACCAGAAGGAGCCGAAGAAUAAAACCUUUCUGAGGUGCGAGGCCAAGAACUAUUCCGGCAGGUUCACGUGCUGGUGGCUUACAACAAUCAGCACAGACCUGACGUUCAGCGUGAAGUCCAGCCGCGGCAGCAGCGACCCCCAGGGGGUGACCUGCGGCGCCGCCACCCUGAGCGCCGAGCGGGUGCGCGGGGACAACAAGGAGUACGAGUACUCCGUGGAGUGCCAGGAAGACAGCGCCUGUCCCGCCGCCGAAGAGAGCCUGCCUAUCGAGGUCAUGGUAGAUGCAGUGCAUAAGCUGAAGUACGAGAACUAUACGAGCAGCUUUUUCAUACGCGACAUCAUCAAGCCCGACCCCCCCAAGAACCUGCAGCUUAAGCCCCUGAAGAAUAGCCGGCAGGUGGAGGUCUCCUGGGAGUACCCCGACACCUGGUCAACGCCCCACAGCUACUUCUCCCUGACCUUUUGUGUCCAAGUCCAGGGAAAGAGCAAGAGGGAGAAGAAAGAUCGGGUGUUCACCGACAAGACCUCCGCCACGGUGAUCUGCAGGAAGAACGCCAGCAUCUCCGUGAGGGCGCAAGACAGGUACUACUCCAGCAGCUGGUCCGAAUGGGCCAGCGUGCCCUGCUCCGGCGGCGGGGGCGGCGGCAGCCGAAACCUACCCGUGGCCACGCCGGAUCCCGGCAUGUUUCCCUGCCUGCACCACAGCCAGAACCUCCUGAGGGCCGUGUCCAACAUGCUGCAGAAGGCCAGGCAGACUCUGGAGUUCUACCCCUGCACGAGCGAGGAGAUCGAUCACGAGGACAUCACCAAGGAUAAGACCAGCACUGUGGAGGCCUGCCUUCCCCUGGAGCUGACCAAGAACGAGAGCUGUCUGAACUCCAGGGAGACCUCAUUCAUCACCAACGGCUCCUGCCUGGCCAGCAGGAAAACCAGCUUCAUGAUGGCCUUGUGUCUCAGCUCCAUCUACGAGGACCUGAAGAUGUAUCAGGUCGAGUUCAAGACAAUGAACGCCAAGCUGCUGAUGGACCCCAAAAGGCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUCAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAGACGGUGCCCCAGAAAAGCUCCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCAGGAUCAGGGCAGUGACCAUCGACCGGGUGAUGUCAUACCUUAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 354 hIL12AB_023UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCAUCAGCAGCUGGU 3′UTR)GAUCUCCUGGUUCAGCCUGGUGUUUCUGGCCUCGCCCCUGGUCGCCAUCUGGGAGCUGAAGAAAGACGUGUACGUCGUCGAACUGGACUGGUACCCCGACGCCCCCGGGGAGAUGGUGGUGCUGACCUGCGACACGCCGGAGGAGGACGGCAUCACCUGGACCCUGGAUCAAAGCAGCGAGGUGCUGGGCAGCGGCAAGACCCUGACCAUCCAAGUGAAGGAAUUCGGCGAUGCCGGCCAGUACACCUGUCACAAAGGGGGCGAGGUGCUCAGCCACAGCCUGCUGCUGCUGCACAAGAAGGAGGAUGGCAUCUGGAGCACCGAUAUCCUGAAGGACCAGAAAGAGCCCAAGAACAAGACGUUCCUGAGGUGCGAGGCCAAGAACUACAGCGGUAGGUUCACGUGUUGGUGGCUGACCACCAUCAGCACCGACCUGACGUUCAGCGUGAAGAGCUCCAGGGGCAGCUCCGACCCACAGGGGGUGACGUGCGGGGCCGCAACCCUCAGCGCCGAAAGGGUGCGGGGGGACAACAAGGAGUACGAAUACUCCGUGGAGUGCCAGGAAGAUUCGGCCUGCCCCGCCGCGGAGGAGAGCCUCCCCAUCGAGGUAAUGGUGGACGCCGUGCAUAAGCUGAAGUACGAGAACUACACCAGCUCGUUCUUCAUCCGAGACAUCAUCAAACCCGACCCGCCCAAAAAUCUGCAGCUCAAGCCCCUGAAGAACUCCAGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGUCCACCCCGCACAGCUACUUCUCCCUGACAUUCUGCGUGCAGGUGCAGGGCAAGAGCAAGCGGGAGAAGAAGGACAGGGUGUUCACCGACAAGACGAGCGCCACCGUGAUCUGCCGAAAGAACGCCAGCAUCUCGGUGCGCGCCCAGGAUAGGUACUAUUCCAGCUCCUGGAGCGAGUGGGCCUCGGUACCCUGCAGCGGCGGCGGGGGCGGCGGCAGUAGGAAUCUGCCCGUGGCUACCCCGGACCCGGGCAUGUUCCCCUGCCUCCACCACAGCCAGAACCUGCUGAGGGCCGUGAGCAACAUGCUGCAGAAGGCCAGACAGACGCUGGAGUUCUACCCCUGCACGAGCGAGGAGAUCGACCACGAGGACAUCACCAAGGAUAAAACUUCCACCGUCGAGGCCUGCCUGCCCUUGGAGCUGACCAAGAAUGAAUCCUGUCUGAACAGCAGGGAGACCUCGUUUAUCACCAAUGGCAGCUGCCUCGCCUCCAGGAAGACCAGCUUCAUGAUGGCCCUCUGUCUGAGCUCCAUCUAUGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCGAAGCUGCUGAUGGACCCCAAGAGGCAGAUCUUCCUGGAUCAGAAUAUGCUGGCGGUGAUCGACGAGCUCAUGCAGGCCCUCAAUUUCAAUAGCGAGACAGUGCCCCAGAAGUCCUCCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGUAUCCUGCUGCACGCCUUCCGGAUCCGGGCCGUCACCAUCGACCGGGUCAUGAGCUACCUCAAUGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 355 hIL12AB_024UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAGCAGCUGGU 3′UTR)GAUCUCCUGGUUCUCCCUGGUGUUCCUGGCCUCGCCCCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUCGUGGAGCUCGACUGGUACCCCGACGCCCCUGGCGAGAUGGUGGUGCUGACCUGCGACACCCCAGAGGAGGAUGGCAUCACCUGGACCCUGGAUCAGUCCUCCGAGGUGCUGGGCUCCGGCAAGACGCUGACCAUCCAAGUGAAGGAGUUCGGUGACGCCGGACAGUAUACCUGCCAUAAGGGCGGCGAGGUCCUGUCCCACAGCCUCCUCCUCCUGCAUAAGAAGGAGGACGGCAUCUGGAGCACCGACAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACCUUUCUGAGGUGCGAGGCCAAGAACUACAGCGGCCGAUUCACCUGCUGGUGGCUCACCACCAUAUCCACCGACCUGACUUUCUCCGUCAAGUCCUCCCGGGGGUCCAGCGACCCCCAGGGAGUGACCUGCGGCGCCGCCACCCUCAGCGCCGAGCGGGUGCGGGGGGACAACAAGGAGUACGAAUACUCCGUCGAGUGCCAGGAGGACUCCGCCUGCCCGGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUCGACGCGGUGCACAAGCUGAAGUACGAGAACUACACCAGCAGUUUCUUCAUCAGGGAUAUCAUCAAGCCAGAUCCCCCGAAGAAUCUGCAACUGAAGCCGCUGAAAAACUCACGACAGGUGGAGGUGAGCUGGGAGUACCCCGACACGUGGAGCACCCCACAUUCCUACUUCAGCCUGACCUUCUGCGUGCAGGUCCAGGGCAAGAGCAAGCGGGAGAAGAAGGACAGGGUGUUCACGGAUAAGACCAGUGCCACCGUGAUCUGCAGGAAGAACGCCUCUAUUAGCGUGAGGGCCCAGGAUCGGUAUUACUCCUCGAGCUGGAGCGAAUGGGCCUCCGUGCCCUGCAGUGGGGGGGGUGGAGGCGGGAGCAGGAACCUGCCCGUAGCAACCCCCGACCCCGGGAUGUUCCCCUGUCUGCACCACUCGCAGAACCUGCUGCGCGCGGUGAGCAACAUGCUCCAAAAAGCCCGUCAGACCUUAGAGUUCUACCCCUGCACCAGCGAAGAAAUCGACCACGAAGACAUCACCAAGGACAAAACCAGCACCGUGGAGGCGUGCCUGCCGCUGGAGCUGACCAAGAACGAGAGCUGCCUCAACUCCAGGGAGACCAGCUUUAUCACCAACGGCUCGUGCCUAGCCAGCCGGAAAACCAGCUUCAUGAUGGCCCUGUGCCUGAGCUCCAUUUACGAGGACCUGAAGAUGUAUCAGGUGGAGUUCAAGACCAUGAAUGCCAAACUCCUGAUGGACCCCAAGAGGCAGAUCUUCCUGGACCAGAACAUGCUCGCGGUGAUCGAUGAGCUGAUGCAGGCCCUGAACUUUAAUAGCGAGACCGUGCCCCAGAAAAGCAGCCUGGAGGAGCCGGACUUCUACAAGACCAAAAUCAAGCUGUGCAUCCUGCUCCACGCCUUCCGCAUCCGGGCCGUGACCAUCGACAGGGUGAUGAGCUACCUGAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 356 hIL12AB_025UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCAUCAGCAGCUGGU 3′UTR)GAUUUCCUGGUUCUCCCUGGUGUUCCUGGCCAGCCCCCUCGUGGCGAUCUGGGAGCUAAAGAAGGACGUGUACGUGGUGGAGCUGGACUGGUACCCGGACGCACCCGGCGAGAUGGUCGUUCUGACCUGCGAUACGCCAGAGGAGGACGGCAUCACCUGGACCCUCGAUCAGAGCAGCGAGGUCCUGGGGAGCGGAAAGACCCUGACCAUCCAGGUCAAGGAGUUCGGCGACGCCGGCCAGUACACCUGCCACAAAGGUGGCGAGGUCCUGAGCCACUCGCUGCUGCUCCUGCAUAAGAAGGAGGACGGAAUCUGGAGCACAGACAUCCUGAAAGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGAGGUGCGAGGCCAAGAACUACAGCGGGCGCUUCACGUGCUGGUGGCUGACCACCAUCAGCACGGACCUCACCUUCUCCGUGAAGAGCAGCCGGGGAUCCAGCGAUCCCCAAGGCGUCACCUGCGGCGCGGCCACCCUGAGCGCGGAGAGGGUCAGGGGCGAUAAUAAGGAGUAUGAGUACAGCGUGGAGUGCCAGGAGGACAGCGCCUGCCCGGCCGCCGAGGAGUCCCUGCCAAUCGAAGUGAUGGUCGACGCCGUGCACAAGCUGAAGUACGAGAACUACACCAGCAGCUUCUUCAUCCGGGAUAUCAUCAAGCCCGAUCCCCCGAAGAACCUGCAGCUGAAGCCCCUCAAGAACAGCCGGCAGGUGGAGGUGAGUUGGGAGUACCCCGACACCUGGUCAACGCCCCACAGCUACUUCUCCCUGACCUUCUGUGUGCAGGUGCAGGGAAAGAGCAAGAGGGAGAAGAAAGACCGGGUCUUCACCGACAAGACCAGCGCCACGGUGAUCUGCAGGAAGAACGCAAGCAUCUCCGUGAGGGCCCAGGACAGGUACUACAGCUCCAGCUGGUCCGAAUGGGCCAGCGUGCCCUGUAGCGGCGGCGGGGGCGGUGGCAGCCGCAACCUCCCAGUGGCCACCCCCGACCCCGGCAUGUUCCCCUGCCUGCACCACAGCCAGAAUCUGCUGAGGGCCGUGAGUAACAUGCUGCAGAAGGCAAGGCAAACCCUCGAAUUCUAUCCCUGCACCUCCGAGGAGAUCGACCACGAGGAUAUCACCAAGGACAAGACCAGCACCGUCGAGGCCUGUCUCCCCCUGGAGCUGACCAAGAAUGAGAGCUGCCUGAACAGCCGGGAGACCAGCUUCAUCACCAACGGGAGCUGCCUGGCCUCCAGGAAGACCUCGUUCAUGAUGGCGCUGUGCCUCUCAAGCAUAUACGAGGAUCUGAAGAUGUACCAGGUGGAGUUUAAGACGAUGAACGCCAAGCUGCUGAUGGACCCGAAGAGGCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUGAUAGACGAGCUCAUGCAGGCCCUGAACUUCAACUCCGAGACCGUGCCGCAGAAGUCAUCCCUCGAGGAGCCCGACUUCUAUAAGACCAAGAUCAAGCUGUGCAUCCUGCUCCACGCCUUCCGGAUAAGGGCCGUGACGAUCGACAGGGUGAUGAGCUACCUUAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 357 hIL12AB_026UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAGCAGCUCGU 3′UTR)GAUCAGCUGGUUCUCCCUGGUGUUUCUCGCCAGCCCCCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGCUGGACUGGUACCCUGACGCCCCGGGGGAGAUGGUCGUGCUGACCUGCGACACCCCCGAAGAGGACGGUAUCACCUGGACCCUGGACCAGUCCAGCGAGGUGCUGGGCAGCGGCAAGACCCUGACUAUUCAAGUCAAGGAGUUCGGAGACGCCGGCCAGUACACCUGCCACAAGGGUGGAGAGGUGUUAUCACACAGCCUGCUGCUGCUGCACAAGAAGGAAGACGGGAUCUGGAGCACCGACAUCCUGAAGGACCAGAAGGAGCCCAAAAACAAGACCUUCCUGCGGUGCGAGGCCAAGAACUAUUCGGGCCGCUUUACGUGCUGGUGGCUGACCACCAUCAGCACUGAUCUCACCUUCAGCGUGAAGUCCUCCCGGGGGUCGUCCGACCCCCAGGGGGUGACCUGCGGGGCCGCCACCCUGUCCGCCGAGAGAGUGAGGGGCGAUAAUAAGGAGUACGAGUACAGCGUUGAGUGCCAGGAAGAUAGCGCCUGUCCCGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUCCACAAGCUGAAGUAUGAGAACUACACCUCAAGCUUCUUCAUCAGGGACAUCAUCAAACCCGAUCCGCCCAAGAAUCUGCAGCUGAAGCCCCUGAAAAAUAGCAGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGUCCACCCCCCAUAGCUAUUUCUCCCUGACGUUCUGCGUGCAGGUGCAAGGGAAGAGCAAGCGGGAGAAGAAGGACCGGGUGUUCACCGACAAGACCUCCGCCACCGUGAUCUGUAGGAAGAACGCGUCGAUCUCGGUCAGGGCCCAGGACAGGUAUUACAGCAGCAGCUGGAGCGAGUGGGCGAGCGUGCCCUGCUCGGGCGGCGGCGGCGGCGGGAGCAGAAAUCUGCCCGUGGCCACCCCAGACCCCGGAAUGUUCCCCUGCCUGCACCAUUCGCAGAACCUCCUGAGGGCCGUGAGCAACAUGCUGCAGAAGGCCCGCCAGACGCUGGAGUUCUACCCCUGCACGAGCGAGGAGAUCGACCACGAAGACAUCACCAAGGACAAAACCAGCACCGUGGAGGCCUGCCUGCCCCUGGAGCUGACCAAAAACGAAUCCUGCCUCAACAGCCGGGAGACCAGCUUCAUCACCAACGGCAGCUGCCUGGCCAGCCGAAAGACCUCCUUCAUGAUGGCCCUCUGCCUGAGCAGCAUCUAUGAGGAUCUGAAGAUGUAUCAGGUGGAGUUCAAGACCAUGAAUGCCAAGCUGCUGAUGGACCCCAAGAGGCAGAUAUUCCUGGACCAGAAUAUGCUGGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAGACCGUCCCCCAGAAGUCCAGCCUGGAGGAGCCGGACUUUUACAAAACGAAGAUCAAGCUGUGCAUACUGCUGCACGCCUUCAGGAUCCGGGCCGUGACAAUCGACAGGGUGAUGUCCUACCUGAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 358 hIL12AB_027UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGUCACCAGCAGCUGGU 3′UTR)GAUCAGCUGGUUCUCCCUGGUGUUCCUGGCCAGCCCCCUGGUGGCCAUCUGGGAGCUCAAGAAGGACGUCUACGUCGUGGAGCUGGAUUGGUACCCCGACGCUCCCGGGGAGAUGGUGGUGCUGACCUGCGACACCCCCGAGGAGGACGGCAUCACCUGGACGCUGGACCAGAGCUCAGAGGUGCUGGGAAGCGGAAAGACACUGACCAUCCAGGUGAAGGAGUUCGGGGAUGCCGGGCAGUAUACCUGCCACAAGGGCGGCGAAGUGCUGAGCCAUUCCCUGCUGCUGCUGCACAAGAAGGAGGACGGCAUAUGGUCCACCGACAUCCUGAAGGAUCAGAAGGAGCCGAAGAAUAAAACCUUCCUGAGGUGCGAGGCCAAGAAUUACAGCGGCCGAUUCACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUGACCUUCAGUGUGAAGUCCUCACGGGGCAGCUCAGAUCCCCAGGGCGUGACCUGCGGGGCCGCGACACUCAGCGCCGAGCGGGUGAGGGGUGAUAACAAGGAGUACGAGUAUUCUGUGGAGUGCCAGGAAGACUCCGCCUGUCCCGCCGCCGAGGAGUCCCUGCCCAUCGAGGUGAUGGUGGACGCCGUGCAUAAACUGAAGUACGAGAACUACACCUCCAGCUUCUUCAUCCGGGAUAUAAUCAAGCCCGACCCUCCGAAAAACCUGCAGCUGAAGCCCCUUAAAAACAGCCGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGAGCACCCCCCAUAGCUAUUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGGAAGUCCAAGCGCGAGAAAAAGGACCGGGUGUUCACCGACAAGACGAGCGCCACCGUGAUCUGCCGGAAGAACGCCAGUAUAAGCGUAAGGGCCCAGGAUAGGUACUACAGCUCCAGCUGGUCGGAGUGGGCCUCCGUGCCCUGUUCCGGCGGCGGGGGGGGUGGCAGCAGGAACCUCCCCGUGGCCACGCCGGACCCCGGCAUGUUCCCGUGCCUGCACCACUCCCAAAACCUCCUGCGGGCCGUCAGCAACAUGCUGCAAAAGGCGCGGCAGACCCUGGAGUUUUACCCCUGUACCUCCGAAGAGAUCGACCACGAGGAUAUCACCAAGGAUAAGACCUCCACCGUGGAGGCCUGUCUCCCCCUGGAGCUGACCAAGAACGAGAGCUGUCUUAACAGCAGAGAGACCUCGUUCAUAACGAACGGCUCCUGCCUCGCUUCCAGGAAGACGUCGUUCAUGAUGGCGCUGUGCCUGUCCAGCAUCUACGAGGACCUGAAGAUGUAUCAGGUCGAGUUCAAAACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGAGGCAGAUCUUCCUGGACCAGAACAUGCUCGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAAACCGUGCCCCAGAAGUCAAGCCUGGAGGAGCCGGACUUCUAUAAGACCAAGAUCAAGCUGUGUAUCCUGCUACACGCUUUUCGUAUCCGGGCCGUGACCAUCGACAGGGUUAUGUCGUACUUGAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 359 hIL12AB_028UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAACAGCUCGU 3′UTR)GAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCAGCCCGCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGCUGGACUGGUACCCCGACGCCCCCGGCGAGAUGGUGGUCCUGACCUGCGACACGCCGGAAGAGGACGGCAUCACCUGGACCCUGGAUCAGUCCAGCGAGGUGCUGGGCUCCGGCAAGACCCUGACCAUUCAGGUGAAGGAGUUCGGCGACGCCGGUCAGUACACCUGCCACAAGGGCGGCGAGGUGCUGAGCCACAGCCUACUGCUCCUGCACAAAAAGGAGGAUGGAAUCUGGUCCACCGACAUCCUCAAGGACCAGAAGGAGCCGAAGAACAAGACGUUCCUCCGGUGCGAGGCCAAGAACUACAGCGGCAGGUUUACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUGACAUUUUCCGUGAAGAGCAGCCGCGGCAGCAGCGAUCCCCAGGGCGUGACCUGCGGGGCGGCCACCCUGUCCGCCGAGCGUGUGAGGGGCGACAACAAGGAGUACGAGUACAGCGUGGAAUGCCAGGAGGACAGCGCCUGUCCCGCCGCCGAGGAGAGCCUGCCAAUCGAGGUCAUGGUGGACGCCGUGCACAAGCUGAAGUACGAGAACUACACGAGCAGCUUCUUCAUCAGGGACAUCAUCAAACCGGACCCGCCCAAGAACCUGCAGCUGAAACCCUUGAAAAACAGCAGGCAGGUGGAAGUGUCUUGGGAGUACCCCGACACCUGGUCCACCCCCCACAGCUACUUUAGCCUGACCUUCUGUGUGCAGGUCCAGGGCAAGUCCAAGAGGGAGAAGAAGGACAGGGUGUUCACCGACAAAACCAGCGCCACCGUGAUCUGCAGGAAGAACGCCUCCAUCAGCGUGCGGGCCCAGGACAGGUAUUACAGCUCGUCGUGGAGCGAGUGGGCCAGCGUGCCCUGCUCCGGGGGAGGCGGCGGCGGAAGCCGGAAUCUGCCCGUGGCCACCCCCGAUCCCGGCAUGUUCCCGUGUCUGCACCACAGCCAGAACCUGCUGCGGGCCGUGAGCAACAUGCUGCAGAAGGCCCGCCAAACCCUGGAGUUCUACCCCUGUACAAGCGAGGAGAUCGACCAUGAGGACAUUACCAAGGACAAGACCAGCACCGUGGAGGCCUGCCUGCCCCUCGAGCUCACAAAGAACGAAUCCUGCCUGAAUAGCCGCGAGACCAGCUUUAUCACGAACGGGUCCUGCCUCGCCAGCCGGAAGACAAGCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAAAUGUACCAAGUGGAGUUCAAAACGAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGCCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUCAUCGACGAGCUCAUGCAGGCCCUGAACUUCAACAGCGAGACCGUGCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUCUACAAGACGAAGAUCAAGCUCUGCAUCCUGCUGCACGCUUUCCGCAUCCGCGCGGUGACCAUCGACCGGGUGAUGAGCUACCUCAACGCCAGUUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 360 hIL12AB_029UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAACAGCUGGU 3′UTR)GAUCAGCUGGUUCAGCCUGGUGUUUCUGGCCUCCCCUCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGCUGGACUGGUACCCUGACGCCCCCGGCGAAAUGGUGGUGCUGACGUGCGACACCCCCGAGGAGGAUGGCAUCACCUGGACCCUGGACCAAAGCAGCGAGGUCCUCGGAAGCGGCAAGACCCUCACUAUCCAAGUGAAGGAGUUCGGGGAUGCGGGCCAGUACACCUGCCACAAGGGCGGCGAGGUGCUGUCUCAUAGCCUGCUGCUCCUGCAUAAGAAGGAAGACGGCAUCUGGAGCACCGACAUACUGAAGGAUCAGAAGGAGCCCAAGAACAAGACCUUCCUGAGGUGCGAGGCCAAGAACUACUCCGGGCGCUUCACCUGUUGGUGGCUGACCACCAUCUCCACCGACCUGACCUUCAGCGUGAAGAGCAGCAGGGGGAGCAGCGACCCCCAGGGGGUGACCUGCGGAGCCGCGACCUUGUCGGCCGAGCGGGUGAGGGGCGACAAUAAGGAGUACGAGUACUCGGUCGAAUGCCAGGAGGACUCCGCCUGCCCCGCCGCCGAGGAGUCCCUCCCCAUCGAAGUGAUGGUGGACGCCGUCCACAAGCUGAAGUACGAGAACUACACCAGCAGCUUCUUCAUACGGGAUAUCAUCAAGCCCGACCCCCCGAAGAACCUGCAGCUGAAACCCUUGAAGAACUCCAGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGUCCACCCCGCACUCAUACUUCAGCCUGACCUUCUGUGUACAGGUCCAGGGCAAGAGCAAGAGGGAAAAGAAGGAUAGGGUGUUCACCGACAAGACCUCCGCCACGGUGAUCUGUCGGAAAAACGCCAGCAUCUCCGUGCGGGCCCAGGACAGGUACUAUUCCAGCAGCUGGAGCGAGUGGGCCUCCGUCCCCUGCUCCGGCGGCGGUGGCGGGGGCAGCAGGAACCUCCCCGUGGCCACCCCCGAUCCCGGGAUGUUCCCAUGCCUGCACCACAGCCAAAACCUGCUGAGGGCCGUCUCCAAUAUGCUGCAGAAGGCGAGGCAGACCCUGGAGUUCUACCCCUGUACCUCCGAGGAGAUCGACCACGAGGAUAUCACCAAGGACAAGACCUCCACGGUCGAGGCGUGCCUGCCCCUGGAGCUCACGAAGAACGAGAGCUGCCUUAACUCCAGGGAAACCUCGUUUAUCACGAACGGCAGCUGCCUGGCGUCACGGAAGACCUCCUUUAUGAUGGCCCUAUGUCUGUCCUCGAUCUACGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGAUCCCAAGAGGCAGAUUUUCCUGGACCAGAACAUGCUGGCCGUGAUUGACGAGCUGAUGCAGGCGCUGAACUUCAACAGCGAGACAGUGCCGCAGAAGAGCUCCCUGGAGGAGCCGGACUUUUACAAGACCAAGAUAAAGCUGUGCAUCCUGCUCCACGCCUUCAGAAUACGGGCCGUCACCAUCGAUAGGGUGAUGUCUUACCUGAACGCCUCCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 361 hIL12AB_030UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAGCAGCUGGU 3′UTR)GAUUAGCUGGUUUAGCCUGGUGUUCCUGGCAAGCCCCCUGGUGGCCAUCUGGGAACUGAAAAAGGACGUGUACGUGGUCGAGCUGGAUUGGUACCCCGACGCCCCCGGCGAAAUGGUGGUGCUGACGUGUGAUACCCCCGAGGAGGACGGGAUCACCUGGACCCUGGAUCAGAGCAGCGAGGUGCUGGGGAGCGGGAAGACCCUGACGAUCCAGGUCAAGGAGUUCGGCGACGCUGGGCAGUACACCUGUCACAAGGGCGGGGAGGUGCUGUCCCACUCCCUGCUGCUCCUGCAUAAGAAAGAGGACGGCAUCUGGUCCACCGACAUCCUCAAGGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGCGGUGUGAGGCGAAGAACUACAGCGGCCGUUUCACCUGCUGGUGGCUGACGACAAUCAGCACCGACUUGACGUUCUCCGUGAAGUCCUCCAGAGGCAGCUCCGACCCCCAAGGGGUGACGUGCGGCGCGGCCACCCUGAGCGCCGAGCGGGUGCGGGGGGACAACAAGGAGUACGAGUACUCCGUGGAGUGCCAGGAGGACAGCGCCUGUCCCGCAGCCGAGGAGUCCCUGCCCAUCGAAGUCAUGGUGGACGCCGUCCACAAGCUGAAGUACGAGAACUACACCAGCAGCUUCUUCAUCCGCGAUAUCAUCAAGCCCGAUCCCCCCAAAAACCUGCAACUGAAGCCGCUGAAGAAUAGCAGGCAGGUGGAGGUGUCCUGGGAGUACCCGGACACCUGGAGCACGCCCCACAGCUAUUUCAGCCUGACCUUUUGCGUGCAGGUCCAGGGGAAGAGCAAGCGGGAGAAGAAGGACCGCGUGUUUACGGACAAAACCAGCGCCACCGUGAUCUGCAGGAAGAACGCCAGCAUCAGCGUGAGGGCCCAGGACAGGUACUACAGCAGCUCCUGGAGCGAGUGGGCCUCCGUGCCCUGUUCCGGAGGCGGCGGGGGCGGUUCCCGGAACCUCCCGGUGGCCACCCCCGACCCGGGCAUGUUCCCGUGCCUGCACCACUCACAGAAUCUGCUGAGGGCCGUGAGCAAUAUGCUGCAGAAGGCAAGGCAGACCCUGGAGUUUUAUCCCUGCACCAGCGAGGAGAUCGACCACGAAGACAUCACCAAGGACAAGACCAGCACAGUGGAGGCCUGCCUGCCCCUGGAACUGACCAAGAACGAGUCCUGUCUGAACUCCCGGGAAACCAGCUUCAUAACCAACGGCUCCUGUCUCGCCAGCAGGAAGACCAGCUUCAUGAUGGCCCUGUGCCUCAGCUCGAUCUACGAGGACCCAAGAUGUACCAGGUUGAGUUCAAGACGAUGAACGCCAAGCUCCUGAUGGACCCCAAGAGGCAGAUCUUCCUGGACCAGAAUAUGCUGGCCGUGAUCGAUGAGUUAAUGCAGGCGCUGAACUUCAACAGCGAGACGGUGCCCCAAAAGUCCUCGCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUCCUGCACGCCUUCCGAAUCCGGGCCGUAACCAUCGACAGGGUGAUGAGCUAUCUCAACGCCUCCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 362 hIL12AB_031UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAGCAGCUCGU 3′UTR)GAUCAGCUGGUUCUCGCUUGUGUUCCUGGCCUCCCCCCUCGUCGCCAUCUGGGAGCUGAAGAAAGACGUGUACGUGGUGGAGCUGGACUGGUAUCCCGACGCCCCGGGGGAGAUGGUGGUGCUGACCUGCGACACCCCGGAAGAGGACGGCAUCACCUGGACGCUCGACCAGUCGUCCGAAGUGCUGGGGUCGGGCAAGACCCUCACCAUCCAGGUGAAGGAGUUCGGAGACGCCGGCCAGUACACCUGUCAUAAGGGGGGGGAGGUGCUGAGCCACAGCCUCCUGCUCCUGCACAAAAAGGAGGACGGCAUCUGGAGCACCGAUAUCCUCAAGGACCAGAAGGAGCCCAAGAACAAGACGUUCCUGAGGUGUGAGGCCAAGAACUACAGCGGGCGGUUCACGUGUUGGUGGCUCACCACCAUCUCCACCGACCUCACCUUCUCCGUGAAGUCAAGCAGGGGCAGCUCCGACCCCCAAGGCGUCACCUGCGGCGCCGCCACCCUGAGCGCCGAGAGGGUCAGGGGGGAUAACAAGGAAUACGAGUACAGUGUGGAGUGCCAAGAGGAUAGCGCCUGUCCCGCCGCCGAAGAGAGCCUGCCCAUCGAAGUGAUGGUGGACGCCGUGCACAAGCUGAAGUACGAGAACUACACCUCCAGCUUCUUCAUCAGGGAUAUCAUCAAGCCCGAUCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCAGGCAGGUGGAGGUGAGCUGGGAGUAUCCCGACACGUGGAGCACCCCGCACAGCUACUUCUCGCUGACCUUCUGCGUGCAGGUGCAAGGGAAGUCCAAGAGGGAGAAGAAGGAUAGGGUGUUCACCGACAAAACGAGCGCCACCGUGAUCUGCCGGAAGAAUGCCAGCAUCUCUGUGAGGGCCCAGGACAGGUACUAUUCCAGCUCCUGGUCGGAGUGGGCCAGCGUGCCCUGUAGCGGCGGGGGCGGGGGCGGCAGCAGGAACCUCCCGGUUGCCACCCCCGACCCCGGCAUGUUUCCGUGCCUGCACCACUCGCAAAACCUGCUGCGCGCGGUCUCCAACAUGCUGCAAAAAGCGCGCCAGACGCUGGAGUUCUACCCCUGCACCAGCGAGGAGAUCGAUCAUGAAGAUAUCACCAAAGACAAGACCUCGACCGUGGAGGCCUGCCUGCCCCUGGAGCUCACCAAGAACGAAAGCUGCCUGAACAGCAGGGAGACAAGCUUCAUCACCAACGGCAGCUGCCUGGCCUCCCGGAAGACCAGCUUCAUGAUGGCCCUGUGCCUGUCCAGCAUCUACGAGGAUCUGAAGAUGUACCAAGUGGAGUUUAAGACCAUGAACGCCAAGCUGUUAAUGGACCCCAAAAGGCAGAUCUUCCUGGAUCAGAACAUGCUGGCCGUCAUCGACGAGCUGAUGCAAGCCCUGAACUUCAACAGCGAGACGGUGCCCCAGAAGAGCAGCCUCGAGGAGCCCGACUUCUAUAAGACCAAGAUAAAGCUGUGCAUUCUGCUGCACGCCUUCAGAAUCAGGGCCGUGACCAUCGAUAGGGUGAUGAGCUACCUGAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 363 hIL12AB_032UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGUCACCAGCAGCUGGU 3′UTR)GAUUUCCUGGUUCAGUCUGGUGUUUCUUGCCAGCCCCCUGGUGGCCAUCUGGGAGCUGAAGAAAGACGUAUACGUCGUGGAGCUGGACUGGUAUCCCGACGCUCCCGGCGAGAUGGUGGUCCUCACCUGCGACACCCCAGAGGAGGACGGCAUCACCUGGACCCUGGACCAGAGCUCCGAGGUCCUGGGCAGCGGUAAGACCCUCACCAUCCAGGUGAAGGAGUUUGGUGAUGCCGGGCAGUAUACCUGCCACAAGGGCGGCGAGGUGCUGUCCCACAGCCUCCUGUUACUGCAUAAGAAGGAGGAUGGCAUCUGGAGCACCGACAUCCUCAAGGACCAGAAAGAGCCCAAGAACAAGACCUUUCUGCGGUGCGAGGCGAAAAAUUACUCCGGCCGGUUCACCUGCUGGUGGCUGACCACCAUCAGCACGGACCUGACGUUCUCCGUGAAGUCGAGCAGGGGGAGCUCCGAUCCCCAGGGCGUGACCUGCGGCGCGGCCACCCUGAGCGCCGAGCGCGUCCGCGGGGACAAUAAGGAAUACGAAUAUAGCGUGGAGUGCCAGGAGGACAGCGCCUGCCCCGCGGCCGAGGAGAGCCUCCCGAUCGAGGUGAUGGUGGAUGCCGUCCACAAGCUCAAAUACGAAAACUACACCAGCAGCUUCUUCAUUAGGGACAUCAUCAAGCCCGACCCCCCCAAAAACCUGCAGCUGAAGCCCCUGAAGAACAGCCGCCAGGUCGAGGUGUCAUGGGAGUACCCAGACACCUGGAGCACCCCCCACUCCUACUUCAGCCUGACCUUCUGCGUCCAGGUGCAGGGAAAGUCCAAACGGGAGAAGAAGGAUAGGGUCUUUACCGAUAAGACGUCGGCCACCGUCAUCUGCAGGAAGAACGCCAGCAUAAGCGUGCGGGCGCAGGAUCGGUACUACAGCUCGAGCUGGUCCGAAUGGGCCUCCGUGCCCUGUAGCGGAGGGGGUGGCGGGGGCAGCAGGAACCUGCCCGUGGCCACCCCGGACCCGGGCAUGUUUCCCUGCCUGCAUCACAGUCAGAACCUGCUGAGGGCCGUGAGCAACAUGCUCCAGAAGGCCCGCCAGACCCUGGAGUUUUACCCCUGCACCAGCGAAGAGAUCGAUCACGAAGACAUCACCAAAGACAAGACCUCCACCGUGGAGGCCUGUCUGCCCCUGGAGCUGACCAAGAACGAGAGCUGUCUGAACAGCAGGGAGACCUCCUUCAUCACCAACGGCUCCUGCCUGGCAUCCCGGAAGACCAGCUUCAUGAUGGCCCUGUGUCUGAGCUCUAUCUACGAGGACCUGAAGAUGUACCAGGUCGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGACAGAUAUUCCUGGACCAGAACAUGCUCGCCGUGAUCGAUGAACUGAUGCAAGCCCUGAACUUCAAUAGCGAGACCGUGCCCCAGAAAAGCAGCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAACUGUGCAUACUGCUGCACGCGUUCAGGAUCCGGGCCGUCACCAUCGACCGGGUGAUGUCCUAUCUGAAUGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 364 hIL12AB_033UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAGCAGCUCGU 3′UTR)GAUUAGCUGGUUUUCGCUGGUGUUCCUGGCCAGCCCUCUCGUGGCCAUCUGGGAGCUGAAAAAAGACGUGUACGUGGUGGAGCUGGACUGGUACCCGGACGCCCCCGGCGAGAUGGUGGUGCUGACGUGCGACACCCCGGAAGAGGACGGCAUCACCUGGACCCUGGACCAGUCAUCCGAGGUCCUGGGCAGCGGCAAGACGCUCACCAUCCAGGUGAAGGAGUUCGGCGACGCCGGCCAGUACACAUGCCAUAAGGGCGGGGAGGUGCUGAGCCACAGCCUGCUCCUCCUGCACAAGAAGGAGGAUGGCAUCUGGUCUACAGACAUCCUGAAGGACCAGAAAGAGCCCAAGAACAAGACCUUCCUCCGGUGCGAGGCCAAGAACUACUCCGGGCGGUUUACUUGUUGGUGGCUGACCACCAUCAGCACCGACCUCACCUUCAGCGUGAAGAGCUCCCGAGGGAGCUCCGACCCCCAGGGGGUCACCUGCGGCGCCGCCACCCUGAGCGCCGAGCGGGUGAGGGGCGACAACAAGGAGUAUGAAUACAGCGUGGAAUGCCAAGAGGACAGCGCCUGUCCCGCGGCCGAGGAAAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUCCACAAACUCAAGUACGAGAACUACACCAGCAGUUUCUUCAUUCGCGACAUCAUCAAGCCGGACCCCCCCAAAAACCUGCAGCUCAAACCCCUGAAGAACAGCAGGCAGGUGGAGGUCAGCUGGGAGUACCCGGACACCUGGAGCACCCCCCAUAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGCAAGAGCAAACGCGAGAAGAAGGACCGGGUGUUUACCGACAAGACCAGCGCCACGGUGAUCUGCCGAAAGAAUGCAAGCAUCUCCGUGAGGGCGCAGGACCGCUACUACUCUAGCAGCUGGAGCGAGUGGGCCAGCGUGCCCUGCAGCGGUGGCGGCGGAGGCGGCAGCCGUAACCUCCCCGUGGCCACCCCCGACCCCGGCAUGUUCCCGUGUCUGCACCACUCCCAGAACCUGCUGAGGGCCGUCAGCAAUAUGCUGCAGAAGGCCCGGCAGACGCUGGAGUUCUACCCCUGCACCUCCGAGGAGAUCGACCAUGAGGACAUUACCAAGGACAAGACGAGCACUGUGGAGGCCUGCCUGCCCCUGGAGCUCACCAAAAACGAGAGCUGCCUGAAUAGCAGGGAGACGUCCUUCAUCACCAACGGCAGCUGUCUGGCCAGCAGGAAGACCAGCUUCAUGAUGGCCCUGUGCCUCUCCUCCAUAUAUGAGGAUCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGAUCCCAAGAGGCAGAUCUUCCUGGACCAGAAUAUGCUGGCCGUGAUUGACGAGCUGAUGCAGGCCCUGAACUUUAAUAGCGAGACCGUCCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUCUAUAAGACCAAGAUCAAGCUGUGCAUACUGCUGCACGCGUUUAGGAUAAGGGCCGUCACCAUCGACAGGGUGAUGAGCUACCUGAAUGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 365 hIL12AB_034UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAACAGCUGGU 3′UTR)GAUCUCCUGGUUCAGCCUGGUGUUCCUCGCCAGCCCCCUGGUGGCCAUCUGGGAGCUGAAGAAAGACGUGUACGUGGUGGAGCUGGACUGGUAUCCCGACGCCCCCGGCGAGAUGGUCGUGCUGACCUGCGACACCCCGGAGGAGGACGGCAUCACCUGGACCCUGGAUCAGUCCUCCGAGGUGCUGGGCAGCGGGAAGACCCUGACCAUCCAGGUGAAAGAGUUCGGAGAUGCCGGCCAGUAUACCUGUCACAAGGGGGGUGAGGUGCUGAGCCAUAGCCUCUUGCUUCUGCACAAGAAGGAGGACGGCAUCUGGUCCACCGACAUCCUCAAGGACCAAAAGGAGCCGAAGAAUAAAACGUUCCUGAGGUGCGAAGCCAAGAACUAUUCCGGACGGUUCACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUCACCUUCUCCGUAAAGUCAAGCAGGGGCAGCUCCGACCCCCAGGGCGUGACCUGCGGAGCCGCCACCCUGAGCGCAGAGAGGGUGAGGGGCGACAACAAGGAGUACGAAUACUCCGUCGAGUGCCAGGAGGACAGCGCCUGCCCCGCCGCCGAGGAAAGUCUGCCCAUCGAGGUGAUGGUGGACGCCGUGCACAAGCUCAAAUACGAGAACUACACCAGCAGCUUCUUCAUCCGGGAUAUCAUCAAGCCCGACCCUCCAAAGAAUCUGCAGCUGAAACCCCUUAAGAACAGCAGGCAGGUGGAGGUCAGCUGGGAGUACCCCGACACCUGGAGCACGCCCCACUCCUACUUUAGCCUGACCUUUUGCGUGCAGGUGCAGGGGAAAAGCAAGCGGGAGAAGAAGGACAGGGUGUUCACCGAUAAGACCUCCGCUACCGUGAUCUGCAGGAAGAACGCCUCAAUCAGCGUGAGGGCCCAGGAUCGGUACUACUCCAGCUCCUGGAGCGAGUGGGCCAGCGUGCCCUGCUCUGGCGGUGGCGGCGGGGGCAGCCGGAACCUGCCGGUGGCCACUCCCGACCCGGGCAUGUUCCCGUGCCUCCACCAUUCCCAGAACCUGCUGCGGGCCGUGUCCAAUAUGCUCCAGAAGGCAAGGCAGACCCUGGAGUUCUACCCCUGCACCAGCGAGGAGAUCGAUCACGAGGACAUCACCAAAGACAAAACCAGCACGGUCGAGGCCUGCCUGCCCCUGGAACUCACCAAGAACGAAAGCUGUCUCAACAGCCGCGAGACCAGCUUCAUAACCAACGGUUCCUGUCUGGCCUCCCGCAAGACCAGCUUUAUGAUGGCCCUCUGUCUGAGCUCCAUCUAUGAAGACCUGAAAAUGUACCAGGUGGAGUUCAAAACCAUGAACGCCAAGCUUCUGAUGGACCCCAAGAGGCAGAUCUUCCUGGAUCAGAACAUGCUGGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUUAACUCCGAGACCGUGCCCCAGAAAAGCAGCCUGGAAGAGCCCGAUUUCUACAAAACGAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCCGGAUCCGUGCGGUGACCAUCGAUAGGGUGAUGAGCUACCUGAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 366 hIL12AB_035UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAACAGCUGGU 3′UTR)AAUCAGCUGGUUCAGCCUGGUUUUCCUCGCGUCGCCCCUGGUGGCCAUCUGGGAGUUAAAGAAGGACGUGUACGUGGUGGAGCUGGAUUGGUACCCCGACGCCCCGGGCGAGAUGGUCGUGCUCACCUGCGAUACCCCCGAGGAGGACGGGAUCACCUGGACCCUGGACCAAUCCAGCGAGGUGCUGGGCAGCGGCAAGACCCUGACCAUACAGGUGAAGGAAUUUGGGGACGCCGGGCAGUACACCUGCCACAAGGGCGGGGAAGUGCUGUCCCACUCCCUCCUGCUGCUGCAUAAGAAGGAGGACGGCAUCUGGAGCACCGACAUCCUGAAGGACCAAAAGGAGCCCAAGAACAAGACCUUCCUGAGGUGCGAGGCCAAAAACUAUUCCGGCCGCUUUACCUGUUGGUGGCUGACCACCAUCUCCACCGAUCUGACCUUCAGCGUGAAGUCGUCUAGGGGCUCCUCCGACCCCCAGGGCGUAACCUGCGGCGCCGCGACCCUGAGCGCCGAGAGGGUGCGGGGCGAUAACAAAGAGUACGAGUACUCGGUGGAGUGCCAGGAGGACAGCGCCUGUCCGGCGGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUCCACAAGCUGAAGUACGAGAACUACACCAGUUCGUUCUUCAUCAGGGACAUCAUCAAGCCGGACCCCCCCAAGAACCUCCAGCUGAAGCCCCUGAAGAACAGCAGGCAGGUGGAAGUGUCCUGGGAGUAUCCCGACACCUGGAGCACCCCCCACAGCUACUUCAGCCUGACCUUUUGCGUGCAGGUGCAGGGCAAAAGCAAGAGGGAAAAGAAGGACCGGGUGUUCACCGAUAAGACGAGCGCCACCGUUAUCUGCAGGAAGAACGCCUCCAUAAGCGUGAGGGCGCAGGACCGUUACUACAGCAGCAGCUGGAGUGAGUGGGCAAGCGUGCCCUGUAGCGGCGGGGGCGGGGGCGGGUCCCGCAACCUCCCCGUCGCCACCCCCGACCCAGGCAUGUUUCCGUGCCUGCACCACAGCCAGAACCUGCUGCGGGCCGUUAGCAACAUGCUGCAGAAGGCCAGGCAGACCCUCGAGUUCUAUCCCUGCACAUCUGAGGAGAUCGACCACGAAGACAUCACUAAGGAUAAGACCUCCACCGUGGAGGCCUGUCUGCCCCUCGAGCUGACCAAGAAUGAAUCCUGCCUGAACAGCCGAGAGACCAGCUUUAUCACCAACGGCUCCUGCCUGGCCAGCAGGAAGACCUCCUUCAUGAUGGCCCUGUGCCUCUCCAGCAUCUACGAGGAUCUGAAGAUGUACCAGGUAGAGUUCAAGACGAUGAACGCCAAGCUCCUGAUGGACCCCAAGAGGCAGAUAUUCCUGGACCAGAACAUGCUGGCGGUGAUCGACGAGCUGAUGCAGGCCCUGAAUUUCAACAGCGAGACGGUGCCACAGAAGUCCAGCCUGGAGGAGCCAGACUUCUACAAGACCAAGAUCAAACUGUGCAUCCUCCUGCACGCGUUCAGGAUCCGCGCCGUCACCAUAGACAGGGUGAUGAGUUAUCUGAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 367 hIL12AB_036UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCAUCAGCAGCUGGU 3′UTR)AAUCAGCUGGUUUAGCCUGGUGUUCCUGGCCAGCCCACUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAACUGGACUGGUACCCCGACGCCCCUGGCGAGAUGGUGGUACUGACCUGUGACACCCCGGAGGAAGACGGUAUCACCUGGACCCUGGAUCAGAGCUCCGAGGUGCUGGGCUCCGGCAAGACACUGACCAUCCAAGUUAAGGAAUUUGGGGACGCCGGCCAGUACACCUGCCACAAGGGGGGCGAGGUGCUGUCCCACUCCCUGCUGCUUCUGCAUAAGAAGGAGGAUGGCAUCUGGUCCACCGACAUACUGAAGGACCAGAAGGAGCCCAAGAAUAAGACCUUCCUGAGAUGCGAGGCCAAGAACUACUCGGGAAGGUUCACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUGACCUUCUCCGUGAAGAGCUCCCGGGGCAGCUCCGACCCCCAGGGCGUAACCUGUGGGGCCGCUACCCUGUCCGCCGAGAGGGUCCGGGGCGACAACAAGGAAUACGAGUACAGCGUGGAGUGCCAGGAGGACUCCGCCUGCCCCGCCGCCGAGGAGUCGCUGCCCAUAGAGGUGAUGGUGGACGCCGUGCACAAGCUCAAGUACGAGAAUUACACCAGCAGCUUCUUUAUCAGGGACAUAAUUAAGCCGGACCCCCCAAAGAAUCUGCAGCUGAAGCCCCUGAAGAAUAGCCGGCAGGUGGAAGUGUCCUGGGAGUACCCCGACACCUGGAGCACCCCCCACUCCUAUUUCUCACUGACAUUCUGCGUGCAGGUGCAAGGGAAAAGCAAGAGGGAGAAGAAGGAUAGGGUGUUCACCGACAAGACAAGCGCCACCGUGAUCUGCCGAAAAAAUGCCAGCAUCAGCGUGAGGGCCCAGGAUCGGUAUUACAGCAGCUCCUGGAGCGAGUGGGCCAGCGUGCCCUGUUCCGGCGGGGGAGGGGGCGGCUCCCGGAACCUGCCGGUGGCCACCCCCGACCCUGGCAUGUUCCCCUGCCUGCAUCACAGCCAGAACCUGCUCCGGGCCGUGUCGAACAUGCUGCAGAAGGCCCGGCAGACCCUCGAGUUUUACCCCUGCACCAGCGAAGAGAUCGACCACGAAGACAUAACCAAGGACAAGACCAGCACGGUGGAGGCCUGCCUGCCCCUGGAGCUUACCAAAAACGAGUCCUGCCUGAACAGCCGGGAAACCAGCUUCAUAACGAACGGGAGCUGCCUGGCCUCCAGGAAGACCAGCUUCAUGAUGGCGCUGUGUCUGUCCAGCAUAUACGAGGAUCUGAAGAUGUAUCAGGUGGAAUUCAAAACUAUGAAUGCCAAGCUCCUGAUGGACCCCAAGAGGCAGAUCUUCCUGGACCAGAACAUGCUAGCCGUGAUCGACGAGCUGAUGCAGGCCCUCAACUUCAACUCGGAGACGGUGCCCCAGAAGUCCAGCCUCGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUACUGCUGCAUGCCUUCAGGAUAAGGGCGGUGACUAUCGACAGGGUCAUGUCCUACCUGAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 368 hIL12AB_037UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAACAACUGGU 3′UTR)GAUCAGCUGGUUCUCCCUGGUGUUCCUGGCCAGCCCCCUGGUGGCCAUCUGGGAGCUCAAAAAAGACGUGUACGUGGUGGAGCUCGAUUGGUACCCAGACGCGCCGGGGGAAAUGGUGGUGCUGACCUGCGACACCCCAGAGGAGGAUGGCAUCACGUGGACGCUGGAUCAGUCCAGCGAGGUGCUGGGGAGCGGCAAGACGCUCACCAUCCAGGUGAAGGAAUUUGGCGACGCGGGCCAGUAUACCUGUCACAAGGGCGGCGAGGUGCUGAGCCACUCCCUGCUGCUGCUGCACAAGAAGGAGGAUGGGAUCUGGUCAACCGAUAUCCUGAAAGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGCGCUGCGAGGCCAAGAACUAUAGCGGCAGGUUCACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUGACCUUCAGCGUGAAAUCCUCCAGGGGCAGCAGCGACCCCCAGGGCGUGACCUGCGGUGCCGCCACGCUCUCCGCCGAGCGAGUGAGGGGUGACAACAAGGAGUACGAGUACAGCGUGGAAUGUCAGGAGGACAGCGCCUGUCCCGCCGCCGAGGAGUCGCUGCCCAUCGAGGUGAUGGUCGACGCGGUGCACAAGCUCAAAUACGAGAAUUACACCAGCAGCUUCUUCAUCAGGGACAUCAUCAAGCCCGACCCCCCCAAGAACCUGCAGCUGAAGCCCUUGAAGAACAGCAGGCAGGUGGAGGUGAGCUGGGAGUACCCGGACACCUGGAGCACCCCCCACUCCUACUUCAGCCUGACGUUCUGUGUGCAGGUGCAGGGGAAGUCCAAGAGGGAGAAGAAGGACCGGGUGUUCACCGACAAGACCAGCGCCACCGUGAUAUGCCGCAAGAACGCGUCCAUCAGCGUUCGCGCCCAGGACCGCUACUACAGCAGCUCCUGGUCCGAAUGGGCCAGCGUGCCCUGCAGCGGUGGAGGGGGCGGGGGCUCCAGGAAUCUGCCGGUGGCCACCCCCGACCCCGGGAUGUUCCCGUGUCUGCAUCACUCCCAGAACCUGCUGCGGGCCGUGAGCAAUAUGCUGCAGAAGGCCAGGCAGACGCUCGAGUUCUACCCCUGCACCUCCGAAGAGAUCGACCAUGAGGACAUCACCAAGGACAAGACCAGCACCGUGGAGGCCUGCCUCCCCCUGGAGCUGACCAAAAACGAGAGCUGCCUGAACUCCAGGGAGACCAGCUUUAUAACCAACGGCAGCUGCCUCGCCUCCAGGAAGACCUCGUUUAUGAUGGCCCUCUGCCUGUCCAGCAUCUACGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCGAAGUUGCUCAUGGACCCCAAGAGGCAGAUCUUCCUGGACCAGAACAUGCUCGCGGUGAUCGACGAGCUGAUGCAAGCCCUGAACUUCAACAGCGAGACCGUGCCCCAGAAGAGCAGCCUGGAAGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCCGGAUCCGGGCCGUGACCAUCGACAGGGUGAUGAGCUACCUCAACGCCUCCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 369 hIL12AB_038UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAGCAGCUCGU 3′UTR)GAUCAGCUGGUUCUCCCUCGUCUUCCUGGCCUCCCCGCUGGUGGCCAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGCUGGACUGGUAUCCCGACGCCCCCGGCGAGAUGGUGGUGCUGACGUGCGACACACCAGAAGAGGACGGGAUCACAUGGACCCUGGAUCAGUCGUCCGAGGUGCUGGGGAGCGGCAAGACCCUCACCAUCCAAGUGAAGGAGUUCGGGGACGCCGGCCAGUACACCUGCCACAAGGGCGGGGAGGUGCUCUCCCAUAGCCUGCUCCUCCUGCACAAAAAGGAGGAUGGCAUCUGGAGCACCGACAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACAUUUCUCAGGUGUGAGGCCAAGAACUAUUCGGGCAGGUUUACCUGUUGGUGGCUCACCACCAUCUCUACCGACCUGACGUUCUCCGUCAAGUCAAGCAGGGGGAGCUCGGACCCCCAGGGGGUGACAUGUGGGGCCGCCACCCUGAGCGCGGAGCGUGUCCGCGGCGACAACAAGGAGUACGAGUAUUCCGUGGAGUGCCAGGAGGACAGCGCCUGCCCCGCCGCCGAGGAGUCCCUGCCCAUAGAGGUGAUGGUGGACGCCGUCCACAAGUUGAAGUACGAAAAUUAUACCUCCUCGUUCUUCAUUAGGGACAUCAUCAAGCCUGACCCCCCGAAGAACCUACAACUCAAGCCCCUCAAGAACUCCCGCCAGGUGGAGGUGUCCUGGGAGUACCCCGACACCUGGUCCACCCCGCACAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUCCAGGGGAAGAGCAAGCGUGAAAAGAAAGACAGGGUGUUCACCGACAAGACGAGCGCCACCGUGAUCUGCAGGAAAAACGCCUCCAUCUCCGUGCGCGCCCAGGACAGGUACUACAGUAGCUCCUGGAGCGAAUGGGCCAGCGUGCCGUGCAGCGGCGGGGGAGGAGGCGGCAGUCGCAACCUGCCCGUGGCCACCCCCGACCCCGGCAUGUUCCCAUGCCUGCACCACAGCCAGAACCUGCUGAGGGCAGUCAGCAAUAUGCUGCAGAAGGCCAGGCAGACCCUGGAGUUUUAUCCCUGCACCAGCGAGGAGAUCGACCACGAGGACAUCACCAAGGACAAGACCUCCACCGUCGAGGCCUGCCUGCCACUGGAGCUGACCAAAAACGAGAGCUGCCUGAACUCCAGGGAGACCUCCUUCAUCACCAACGGGAGCUGCCUGGCCAGCCGGAAGACCAGCUUCAUGAUGGCGCUGUGCCUCAGCAGCAUCUACGAGGAUCUCAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCGAAGCUGCUGAUGGACCCCAAGCGGCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUGAUUGACGAGCUCAUGCAGGCCCUGAACUUCAAUAGCGAGACCGUCCCCCAAAAGAGCAGCCUGGAGGAACCCGACUUCUACAAAACGAAGAUCAAGCUCUGCAUCCUGCUGCACGCCUUCCGGAUCCGGGCCGUGACCAUCGAUCGUGUGAUGAGCUACCUGAACGCCUCGUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 370 hIL12AB_039UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCACCAGCAGCUCGU 3′UTR)CAUCUCCUGGUUUAGCCUGGUGUUUCUGGCCUCCCCCCUGGUCGCCAUCUGGGAGCUGAAGAAAGACGUGUACGUGGUGGAGCUGGACUGGUACCCGGACGCUCCCGGGGAGAUGGUGGUGCUGACCUGCGACACCCCCGAGGAGGACGGCAUCACCUGGACCCUGGACCAGAGCUCCGAGGUGCUGGGGAGCGGCAAGACCCUGACCAUUCAGGUGAAAGAGUUCGGCGACGCCGGCCAAUAUACCUGCCACAAGGGGGGGGAGGUCCUGUCGCAUUCCCUGCUGCUGCUUCACAAAAAGGAGGAUGGCAUCUGGAGCACCGACAUCCUGAAGGACCAGAAAGAACCCAAGAACAAGACGUUCCUGCGCUGCGAGGCCAAGAACUACAGCGGCCGGUUCACCUGUUGGUGGCUGACCACCAUCUCCACCGACCUGACUUUCUCGGUGAAGAGCAGCCGCGGGAGCAGCGACCCCCAGGGAGUGACCUGCGGCGCCGCCACCCUGAGCGCCGAAAGGGUGAGGGGCGACAAUAAAGAGUACGAGUAUUCCGUGGAGUGCCAGGAGGACAGCGCCUGUCCCGCCGCCGAGGAGUCCCUGCCUAUCGAGGUGAUGGUCGACGCGGUGCACAAGCUCAAGUACGAAAACUACACCAGCAGCUUUUUCAUCAGGGAUAUCAUCAAACCAGACCCCCCCAAGAACCUGCAGCUGAAGCCCCUGAAAAACAGCAGGCAGGUGGAAGUGAGCUGGGAAUACCCCGAUACCUGGUCCACCCCCCACAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGGAAGUCCAAGCGGGAGAAGAAAGAUCGGGUGUUCACGGACAAGACCAGCGCCACCGUGAUUUGCAGGAAAAACGCCAGCAUCUCCGUGAGGGCUCAGGACAGGUACUACAGCUCCAGCUGGAGCGAGUGGGCCUCCGUGCCUUGCAGCGGGGGAGGAGGCGGCGGCAGCAGGAAUCUGCCCGUCGCAACCCCCGACCCCGGCAUGUUCCCCUGCCUGCACCACAGCCAGAAUCUGCUGCGAGCCGUGAGCAACAUGCUCCAGAAGGCCCGGCAGACGCUGGAGUUCUACCCCUGCACCUCCGAGGAGAUCGACCACGAGGACAUCACCAAGGAUAAGACGAGCACCGUCGAGGCCUGUCUCCCCCUGGAGCUCACCAAGAACGAGUCCUGCCUGAAUAGCAGGGAGACGUCCUUCAUAACCAACGGCAGCUGUCUGGCGUCCAGGAAGACCAGCUUCAUGAUGGCCCUCUGCCUGAGCUCCAUCUACGAGGACCUCAAGAUGUACCAGGUCGAGUUCAAGACCAUGAACGCAAAACUGCUCAUGGAUCCAAAGAGGCAGAUCUUUCUGGACCAGAACAUGCUGGCCGUGAUCGAUGAACUCAUGCAGGCCCUGAAUUUCAAUUCCGAGACCGUGCCCCAGAAGAGCUCCCUGGAGGAACCCGACUUCUACAAAACAAAGAUCAAGCUGUGUAUCCUCCUGCACGCCUUCCGGAUCAGGGCCGUCACCAUUGACCGGGUGAUGUCCUACCUGAACGCCAGCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 371 hIL12AB_040UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAAAUAAGA (5′UTR ORFGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGUGCCAUCAGCAGCUGGU 3′UTR)GAUCAGCUGGUUCAGCCUCGUGUUCCUCGCCAGCCCCCUCGUGGCCAUCUGGGAGCUGAAAAAGGACGUGUACGUGGUGGAGCUGGACUGGUAUCCCGACGCCCCGGGCGAGAUGGUGGUGCUGACCUGCGACACCCCCGAGGAGGACGGCAUUACCUGGACACUGGACCAGAGCAGCGAGGUCCUGGGCAGCGGGAAGACCCUGACAAUUCAGGUGAAGGAGUUCGGCGACGCCGGACAGUACACGUGCCACAAGGGGGGGGAGGUGCUGUCCCACAGCCUCCUCCUGCUGCACAAGAAGGAGGAUGGCAUCUGGAGCACCGACAUCCUGAAGGAUCAGAAGGAGCCCAAGAACAAGACCUUUCUGAGAUGCGAGGCCAAGAAUUACAGCGGCCGUUUCACCUGCUGGUGGCUCACCACCAUCAGCACCGACCUGACCUUCAGCGUGAAAUCCUCCAGGGGCUCCUCCGACCCGCAGGGAGUGACCUGCGGCGCCGCCACACUGAGCGCCGAGCGGGUCAGAGGGGACAACAAGGAGUACGAGUACAGCGUUGAGUGCCAGGAGGACAGCGCCUGUCCCGCGGCCGAGGAAUCCCUGCCCAUCGAGGUGAUGGUGGACGCAGUGCACAAGCUGAAGUACGAGAACUAUACCUCGAGCUUCUUCAUCCGGGAUAUCAUUAAGCCCGAUCCCCCGAAGAACCUGCAGCUCAAACCCCUGAAGAACAGCAGGCAGGUGGAGGUCUCCUGGGAGUACCCCGACACAUGGUCCACCCCCCAUUCCUAUUUCUCCCUGACCUUUUGCGUGCAGGUGCAGGGCAAGAGCAAGAGGGAGAAAAAGGACAGGGUGUUCACCGACAAGACCUCCGCCACCGUGAUCUGCCGUAAGAACGCUAGCAUCAGCGUCAGGGCCCAGGACAGGUACUAUAGCAGCUCCUGGUCCGAGUGGGCCAGCGUCCCGUGCAGCGGCGGGGGCGGUGGAGGCUCCCGGAACCUCCCCGUGGCCACCCCGGACCCCGGGAUGUUUCCCUGCCUGCAUCACAGCCAGAACCUGCUGAGGGCCGUGUCCAACAUGCUGCAGAAGGCCAGGCAGACACUCGAGUUUUACCCCUGCACCAGCGAGGAGAUCGACCACGAAGACAUCACCAAGGACAAGACCUCCACCGUGGAGGCAUGCCUGCCCCUGGAGCUGACCAAAAACGAAAGCUGUCUGAACUCCAGGGAGACCUCCUUUAUCACGAACGGCUCAUGCCUGGCCUCCAGAAAGACCAGCUUCAUGAUGGCCCUGUGCCUGAGCUCCAUCUACGAGGACUUGAAAAUGUACCAGGUCGAGUUCAAGACCAUGAACGCCAAGCUGCUCAUGGACCCCAAAAGGCAGAUCUUUCUGGACCAGAAUAUGCUGGCCGUGAUCGACGAGCUCAUGCAAGCCCUGAAUUUCAACAGCGAGACCGUGCCCCAGAAGUCCUCCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUACUCCUGCACGCGUUUAGGAUCAGGGCGGUGACCAUCGAUAGGGUGAUGAGCUACCUGAAUGCCUCCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCCAAACACCAUUGUCACACUCCAGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC 372 IL12B signalAUGUGCCACCAGCAGCUGGUGAUCAGCUGGUUCAGCCUGGUGUUCCUGGCCAGCCC peptideCCUGGUGGCC nucleotide sequence 373 IL-12BAUCUGGGAGCUGAAGAAGGACGUGUACGUGGUGGAGUUGGAUUGGUACCCCGACGC nucleotideCCCCGGCGAGAUGGUGGUGCUGACCUGCGACACCCCCGAGGAGGACGGCAUCACCU sequenceGGACCCUGGACCAGAGCAGCGAGGUGCUGGGCAGCGGCAAGACCCUGACCAUCCAGGUGAAGGAGUUCGGCGACGCCGGCCAGUACACCUGCCACAAGGGCGGCGAGGUGCUGAGCCACAGCCUGCUGCUGCUGCACAAGAAGGAGGACGGCAUCUGGAGCACCGACAUCCUGAAGGACCAGAAGGAGCCCAAGAACAAGACCUUCCUGAGAUGCGAGGCCAAGAACUACAGCGGCAGAUUCACCUGCUGGUGGCUGACCACCAUCAGCACCGACCUGACCUUCAGCGUGAAGAGCAGCAGAGGCAGCAGCGACCCCCAGGGCGUGACCUGCGGCGCCGCCACCCUGAGCGCCGAGAGAGUGAGAGGCGACAACAAGGAGUACGAGUACAGCGUGGAGUGCCAGGAAGAUAGCGCCUGCCCCGCCGCCGAGGAGAGCCUGCCCAUCGAGGUGAUGGUGGACGCCGUGCACAAGCUGAAGUACGAGAACUACACCAGCAGCUUCUUCAUCAGAGAUAUCAUCAAGCCCGACCCCCCGAAGAACCUGCAGCUGAAGCCCCUGAAGAACAGCCGGCAGGUGGAGGUGAGCUGGGAGUACCCCGACACCUGGAGCACCCCCCACAGCUACUUCAGCCUGACCUUCUGCGUGCAGGUGCAGGGCAAGAGCAAGAGAGAGAAGAAAGAUAGAGUGUUCACCGACAAGACCAGCGCCACCGUGAUCUGCAGAAAGAACGCCAGCAUCAGCGUGAGAGCCCAAGAUAGAUACUACAGCAGCAGCUGGAGCGAGUGGGCCAGCGUGCCCUGCAGC 374 GGGGGGS GGCGGCGGCGGCGGCGGCAGC nucleotidesequence 375 IL-12AAGAAACCUGCCCGUGGCCACCCCCGACCCCGGCAUGUUCCCCUGCCUGCACCACAG nucleotideCCAGAACCUGCUGAGAGCCGUGAGCAACAUGCUGCAGAAGGCCCGGCAGACCCUGG sequenceAGUUCUACCCCUGCACCAGCGAGGAGAUCGACCACGAAGAUAUCACCAAAGAUAAGACCAGCACCGUGGAGGCCUGCCUGCCCCUGGAGCUGACCAAGAACGAGAGCUGCCUGAACAGCAGAGAGACCAGCUUCAUCACCAACGGCAGCUGCCUGGCCAGCAGAAAGACCAGCUUCAUGAUGGCCCUGUGCCUGAGCAGCAUCUACGAGGACCUGAAGAUGUACCAGGUGGAGUUCAAGACCAUGAACGCCAAGCUGCUGAUGGACCCCAAGCGGCAGAUCUUCCUGGACCAGAACAUGCUGGCCGUGAUCGACGAGCUGAUGCAGGCCCUGAACUUCAACAGCGAGACCGUGCCCCAGAAGAGCAGCCUGGAGGAGCCCGACUUCUACAAGACCAAGAUCAAGCUGUGCAUCCUGCUGCACGCCUUCAGAAUCAGAGCCGUGACCAUCGACAGAGUGAUGAGCUACCUGAACGCCAGC 376 LinkerUCUGGUGGCGGAUCAGGCGGCGGCGGUUCAGGAGGCGGUGGAAGUGGAGGUGGCGG nucleotideGUCUGGCGGAGGUUCACUGCAG sequence 377 Human CD8AUCUACAUCUGGGCUCCACUGGCCGGCACCUGCGGCGUGCUGCUGCUGAGCCUGGU transmembraneGAUCACCCUGUACUGCUAC nucleotide sequence 378 Human CD80CUGCUGCCCAGCUGGGCCAUCACCCUGAUCAGCGUGAACGGCAUCUUCGUGAUCUG transmembraneCUGCCUG domain nucleotide sequence 379 Human CD80ACCUACUGCUUCGCCCCUCGAUGCAGAGAGAGAAGAAGAAACGAGAGACUGAGAAG intracellularAGAGAGCGUGCGACCCGUG domain nucleotide sequence

1-113. (canceled)
 114. A method of treating cancer in a subject in needthereof, comprising administering a lipid nanoparticle (LNP) comprisinga messenger RNA (mRNA) comprising an open reading frame (ORF) encoding ahuman interleukin-12 (IL-12) polypeptide operably linked to a membranedomain, optionally via a linker, wherein the human IL-12 polypeptidecomprises an IL-12 p40 subunit (IL-12B) polypeptide operably linked toan IL-12 p35 subunit (IL-12A) polypeptide, wherein the membrane domaincomprises a transmembrane domain comprising a CD80 transmembrane domain,a PDGFR transmembrane domain, or a CD8 transmembrane domain, therebyreducing the size of the tumor or inhibiting growth of the tumor in thesubject.
 115. The method of claim 114, wherein the membrane domain isoperably linked to the IL-12A polypeptide by a peptide linker, orwherein the membrane domain is operably linked to the IL-12B polypeptideby a peptide linker.
 116. The method of claim 114, wherein the IL-12Bpolypeptide is operably linked to the IL-12A polypeptide by a peptidelinker.
 117. The method of claim 114, wherein (i) the IL-12B polypeptideis located at the 5′ terminus of the IL-12A polypeptide, or the 5′terminus of the peptide linker, or (ii) the IL-12A polypeptide islocated at the 5′ terminus of the IL-12B polypeptide, or the 5′ terminusof the peptide linker.
 118. The method of claim 114, wherein the IL-12Bpolypeptide comprises amino acids 23 to 238 of SEQ ID NO:
 48. 119. Themethod of claim 114, wherein the IL-12A polypeptide comprises aminoacids 336 to 532 of SEQ ID NO:
 48. 120. The method of claim 114, whereinthe mRNA comprises a 3′ untranslated region (UTR) comprising a microRNAbinding site.
 121. The method of claim 120, wherein the microRNA bindingsite is a miR-122 binding site.
 122. The method of claim 121, whereinthe miR-122 binding site is a miR-122-3p or a miR-122-5p binding site123. The method of claim 114, wherein the mRNA comprises at least onechemical modification.
 124. The method of claim 123, wherein thechemical modification is selected from the group consisting ofpseudouridine or a psuedouridine analog.
 125. The method of claim 123,wherein the chemical modification is N1-methylpseudouridine.
 126. Themethod of claim 123, wherein the mRNA is fully modified withN1-methylpseudouridine.
 127. The method of claim 114, wherein the CD80transmembrane domain comprises the amino acid sequence set forth in SEQID NO: 103, the PDGFR transmembrane domain comprises a PDGFR betatransmembrane domain comprising the amino acid sequence set forth in SEQID NO: 102, and the CD8 transmembrane domain comprises the amino acidsequence set forth in SEQ ID NO:
 101. 128. The method of claim 114,wherein the membrane domain comprises an intracellular domain.
 129. Themethod of claim 128, wherein the intracellular domain is a CD80intracellular domain or a PDGFR intracellular domain.
 130. The method ofclaim 129, wherein the CD80 intracellular domain comprises the aminoacid sequence set forth in SEQ ID NO: 225 and the PDGFR intracellulardomain comprises an amino acid sequence selected from SEQ ID NOs:226-228
 131. The method of claim 114, wherein the IL-12 polypeptide isoperably linked to the membrane domain via a peptide linker.
 132. Themethod of claim 114, wherein the ORF encodes the amino acid sequence setforth in any one of SEQ ID NOs: 241, 243, 249, 253 and
 255. 133. Themethod of claim 114, comprising administering a second mRNA encoding asecond polypeptide.
 134. The method of claim 133, wherein the LNPcomprises the second mRNA.
 135. The method of claim 114, whereintreating comprising reducing the size of a tumor or inhibiting growth ofa tumor in the subject.
 136. The method of claim 114, wherein the LNP isadministered intratumorally.
 137. The method of claim 114, wherein theLNP comprises an ionizable amino lipid, a phospholipid, a sterol, and aPEG-modified lipid.
 138. The method of claim 137, wherein the LNPcomprises a molar ratio of about 20-60% ionizable amino lipid: 5-25%phospholipid: 25-55% sterol: 0.5-15% PEG-modified lipid.
 139. A methodof treating cancer in a subject in need thereof, comprisingadministering a LNP comprising a mRNA comprising a 5′ untranslatedregion (UTR), an ORF, and a 3′ UTR, wherein the ORF comprises anucleotide sequence encoding from 5′ to 3′:5′-[IL-12B]-[L1]-[IL-12A]-[L2]-[MD]-3′, wherein IL-12B is a human IL-12p40 subunit polypeptide comprising amino acids 23 to 328 of SEQ ID NO:48, L1 is a first peptide linker, IL-12A is a human IL-12 p35 subunitpolypeptide comprising amino acids 336 to 532 of SEQ ID NO: 48, L2 is asecond peptide linker, and MD is a membrane domain comprising a CD80transmembrane domain and a CD80 intracellular domain.
 140. The method ofclaim 139, wherein the CD80 transmembrane domain comprises the aminoacid sequence set forth in SEQ ID NO: 103 and the CD80 intracellulardomain comprises the amino acid sequence set forth in SEQ ID NO: 225.141. The method of claim 139, wherein the 3′UTR comprises a microRNAbinding site.
 142. The method of claim 141, wherein the microRNA bindingsite is a miR-122 binding site.
 143. The method of claim 142, whereinthe miR-122 binding site is a miR-122-3p or a miR-122-5p binding site.144. The method of claim 139, wherein the mRNA comprises at least onechemical modification.
 145. The method of claim 144, wherein thechemical modification is selected from the group consisting ofpseudouridine or a psuedouridine analog.
 146. The method of claim 144,wherein the chemical modification is N1-methylpseudouridine.
 147. Themethod of claim 144, wherein the mRNA is fully modified withN1-methylpseudouridine.
 148. The method of claim 139, wherein the ORFencodes a signal peptide.
 149. The method of claim 148, wherein thesignal peptide is an IL-12B signal peptide comprising the amino acidsequence set forth in amino acids 1 to 22 of SEQ ID NO:
 48. 150. Themethod of claim 139, wherein [L1] and [L2] are each a Gly/Ser linker.151. The method of claim 139, wherein the ORF encodes the amino acidsequence set forth in SEQ ID NO:
 249. 152. The method of claim 139,comprising administering a second mRNA encoding a second polypeptide.153. The method of claim 152, wherein the LNP comprises the second mRNA.154. The method of claim 139, wherein treating comprising reducing thesize of a tumor or inhibiting growth of a tumor in the subject.
 155. Themethod of claim 139, wherein the LNP is administered intratumorally.156. The method of claim 139, wherein the LNP comprises an ionizableamino lipid, a phospholipid, a sterol, and a PEG-modified lipid. 157.The method of claim 156 wherein the LNP comprises a molar ratio of about20-60% ionizable amino lipid: 5-25% phospholipid: 25-55% sterol: 0.5-15%PEG-modified lipid.